U.S. patent application number 15/587884 was filed with the patent office on 2017-08-31 for tetracyclic anthraquinone derivatives.
The applicant listed for this patent is TIANJIN HEMAY ONCOLOGY PHARMACEUTICAL CO., LTD.. Invention is credited to Aihong Huo, Zhenzhong Li, Hesheng Zhang.
Application Number | 20170247403 15/587884 |
Document ID | / |
Family ID | 50487501 |
Filed Date | 2017-08-31 |
United States Patent
Application |
20170247403 |
Kind Code |
A1 |
Zhang; Hesheng ; et
al. |
August 31, 2017 |
TETRACYCLIC ANTHRAQUINONE DERIVATIVES
Abstract
Disclosed are a compound represented by formula (I) and a
pharmaceutically acceptable salt thereof, ##STR00001## wherein
R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5, W, n are defined as in
the present application.
Inventors: |
Zhang; Hesheng; (Tianjin,
CN) ; Huo; Aihong; (Tianjin, CN) ; Li;
Zhenzhong; (Tianjin, CN) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
TIANJIN HEMAY ONCOLOGY PHARMACEUTICAL CO., LTD. |
TIANJIN |
|
CN |
|
|
Family ID: |
50487501 |
Appl. No.: |
15/587884 |
Filed: |
May 5, 2017 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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14383523 |
Sep 5, 2014 |
9670242 |
|
|
PCT/CN2013/000233 |
Mar 6, 2013 |
|
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15587884 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
C07H 19/044 20130101;
A61P 43/00 20180101; C07H 1/00 20130101; A61P 35/00 20180101; A61P
35/02 20180101 |
International
Class: |
C07H 19/044 20060101
C07H019/044; C07H 1/00 20060101 C07H001/00 |
Foreign Application Data
Date |
Code |
Application Number |
Mar 6, 2013 |
CN |
201210055298.7 |
Mar 6, 2013 |
CN |
PCT/CN2013/000233 |
Claims
1.-22. (canceled)
23. A method for treating-tumor and/or cancer, comprising
administering to a subject in need thereof a therapeutically
effective amount of a compound represented by formula (I) or a
pharmaceutically acceptable salt thereof, or a formulation
comprising a compound represented by formula (I) or a
pharmaceutically acceptable salt thereof, and a pharmaceutically
acceptable carrier, ##STR00245## wherein: R.sup.1 is selected from
the group consisting of H, optionally substituted alkyl, and
optionally substituted alkoxy; R.sup.2 is selected from the group
consisting of H, optionally substituted aryl, optionally
substituted heteroaryl, optionally substituted
(alkyleneoxy).sub.malkyl, optionally substituted heterocyclyl,
optionally substituted alkyl, and optionally substituted
--S(.dbd.O).sub.2R; R.sup.3 is selected from the group consisting
of H, optionally substituted aryl, optionally substituted
heteroaryl, optionally substituted (alkyleneoxy).sub.malkyl,
optionally substituted heterocyclyl, optionally substituted alkyl,
and optionally substituted --S(.dbd.O).sub.2R; or NR.sup.2R.sup.3
represents optionally substituted heterocyclyl; wherein R comprises
optionally substituted alkyl, optionally substituted
cyclohydrocarbyl, optionally substituted heterocyclyl, optionally
substituted aryl, or optionally substituted heteroaryl; m is
selected from the group consisting of 0, 23, 2, 3, 4, 5, 6, 7, 8,
9, 10, and 11; W is selected from the group consisting of O and NH;
R.sup.4 is selected from the group consisting of H, F, and
optionally substituted alkyl; R.sup.5 is selected from the group
consisting of H, F, optionally substituted alkyl and OR.sup.6,
wherein R.sup.6 is selected from the group consisting of H and
tetrahydropyran-2-yl; and n is selected from the group consisting
of 23, 2 and 3; wherein the tumor and/or cancer is selected from
the group consisting of gastric cancer, intestine cancer, head and
neck cancer, cervical cancer, brain glioma, various leukemia,
lymphoma, and multiple bone marrow cancer.
24. The method of claim 23, wherein R.sup.1 is selected from the
group consisting of H and OCH.sub.3.
25. The method of claim 23, wherein W is O.
26. The method of claim 23, wherein R.sup.4 is CH.sub.3.
27. The method of claim 23, wherein R.sup.5 is selected from the
group consisting of OH and (tetrahydropyran-2-yl)oxy.
28. The method of claim 23, wherein R.sup.2 is selected from the
group consisting of H, methyl, ethyl, (morpholinylmethyl)phenyl,
4-((morpholin-1-yl)methyl)phenyl, (dimethylaminomethyl)phenyl,
4-((dimethylamino)methyl)phenyl, 2-(2-(dimethylamino)ethoxy)ethyl,
morpholin-1-yl, piperidin-1-yl, tetrahydropyrrol-1-yl,
4-(2-hydroxyethyl)piperazin-1-yl, (4-methylpiperazin)-1-yl,
(4-ethylpiperazin)-1-yl, 2-(tetrahydropyrrol-1-yl)ethyl,
3-(tetrahydropyrrol-1-yl)propyl, (2-(morpholin-1-yl)pyridin)-4-yl,
(2-(morpholin-1-yl)pyridin)-5-yl, pyridin-2-yl, pyridin-3-yl,
pyridin-4-yl, (pyridin-4-yl)methyl, (pyridin-3-yl)methyl,
(pyridin-2-yl)methyl, 2-(pyridin-4-yl)ethyl, 2-(pyridin-3-yl)ethyl,
2-(pyridin-2-yl)ethyl, 2-(pyridin-4-yl)propyl,
2-(pyridin-3-yl)propyl, 2-(pyridin-2-yl)propyl,
2-((4-sulfamido)phenyl)ethyl,
(3-(dimethylamino)propyl)piperazin-1-yl,
3-((4-sulfamido)phenyl)propyl, 3-((4-methyl)piperazin-1-yl)propyl,
3-((4-ethyl)piperazin-1-yl)propyl,
3-((4-propyl)piperazin-1-yl)propyl,
2-((4-methyl)piperazin-1-yl)ethyl,
2-((4-ethyl)piperazin-1-yl)ethyl,
2-((4-propyl)piperazin-1-yl)ethyl, 2-(dimethylamino)ethyl,
2-(diethylamino)ethyl, 2-(dipropylamino)ethyl,
2-(piperidin-1-yl)ethyl, 2-(morpholin-1-yl)ethyl,
2-(tetrahydropyrrol-1-yl)ethyl, 3-(dimethylamino)propyl,
3-(diethylamino)propyl, 3-(dipropylamino)propyl,
3-(piperidin-1-yl)propyl, 3-(morpholin-1-yl)propyl,
3-(tetrahydropyrrol-1-yl)propyl, 4-(dimethylamino)butyl,
4-(diethylamino)butyl, 4-(dipropylamino)butyl,
4-(piperidin-1-yl)butyl, 4-(morpholin-1-yl)butyl,
4-(tetrahydropyrrol-1-yl)butyl,
2-(2-(2-(2-(dimethylamino)ethoxy)ethoxy)ethoxy)ethyl,
2-(2-(2-(2-(diethylamino)ethoxy)ethoxy)ethoxy)ethyl,
2-(2-(2-(2-(dipropylamino)ethoxy)ethoxy)ethoxy)ethyl,
2-(2-(2-(2-(piperidin-1-yl)ethoxy)ethoxy)ethoxy)ethyl,
2-(2-(2-(2-(morpholin-1-yl)ethoxy)ethoxy)ethoxy)ethyl,
2-(2-(2-(2-(tetrahydropyrrol-1-yl)ethoxy)ethoxy)ethoxy)ethyl,
2-(2-(2-(dimethylamino)ethoxy)ethoxy)ethyl,
2-(2-(2-(diethylamino)ethoxy)ethoxy)ethyl,
2-(2-(2-(dipropylamino)ethoxy)ethoxy)ethyl,
2-(2-(2-(piperidin-1-yl)ethoxy)ethoxy)ethyl,
2-(2-(2-(morpholin-1-yl)ethoxy)ethoxy)ethyl,
2-(2-(2-(tetrahydropyrrol-1-yl)ethoxy)ethoxy)ethyl, 6-purinyl,
mesyl, benzenesulfonyl, pyrazin-2-yl, pyrimidin-2-yl,
2-hydroxyethyl, 2-(2-hydroxyethoxy)ethyl,
2-(2-(2-(2-(2-(2-(2-(2-(2-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)ethoxy)eth-
oxy)ethoxy)ethoxy)ethoxy)ethoxy)ethoxy)ethoxy)ethyl,
2-(2-(2-(2-(2-(2-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)ethoxy)ethoxy)ethox-
y)ethoxy)ethoxy)ethyl,
2-(2-(2-(2-(2-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)ethoxy)ethoxy)ethoxy)e-
thoxy)ethyl,
2-(2-(2-(2-(2-(2-(2-(2-(2-methoxyethoxy)ethoxy)ethoxy)ethoxy)ethoxy)ethox-
y)ethoxy)ethyl, and
2-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)ethyl.
29. The method of claim 23, wherein R.sup.3 is selected from the
group consisting of H, methyl, ethyl, (morpholinylmethyl)phenyl,
4-((morpholin-1-yl)methyl)phenyl, (dimethylaminomethyl)phenyl,
4-((dimethylamino)methyl)phenyl, 2-(2-(dimethylamino)ethoxy)ethyl,
morpholin-1-yl, piperidin-1-yl, tetrahydropyrrol-1-yl,
4-(2-hydroxyethyl)piperazin-1-yl, (4-methylpiperazin)-1-yl,
(4-ethylpiperazin)-1-yl, 2-(tetrahydropyrrol-1-yl)ethyl,
3-(tetrahydropyrrol-1-yl)propyl, (2-(morpholin-1-yl)pyridin)-4-yl,
(2-(morpholin-1-yl)pyridin)-5-yl, pyridin-2-yl, pyridin-3-yl,
pyridin-4-yl, (pyridin-4-yl)methyl, (pyridin-3-yl)methyl,
(pyridin-2-yl)methyl, 2-(pyridin-4-yl)ethyl, 2-(pyridin-3-yl)ethyl,
2-(pyridin-2-yl)ethyl, 2-(pyridin-4-yl)propyl,
2-(pyridin-3-yl)propyl, 2-(pyridin-2-yl)propyl,
2-((4-sulfamido)phenyl)ethyl,
(3-(dimethylamino)propyl)piperazin-1-yl,
3-((4-sulfamido)phenyl)propyl, 3-((4-methyl)piperazin-1-yl)propyl,
3-((4-ethyl)piperazin-1-yl)propyl,
3-((4-propyl)piperazin-1-yl)propyl,
2-((4-methyl)piperazin-1-yl)ethyl,
2-((4-ethyl)piperazin-1-yl)ethyl,
2-((4-propyl)piperazin-1-yl)ethyl, 2-(dimethylamino)ethyl,
2-(diethylamino)ethyl, 2-(dipropylamino)ethyl,
2-(piperidin-1-yl)ethyl, 2-(morpholin-1-yl)ethyl,
2-(tetrahydropyrrol-1-yl)ethyl, 3-(dimethylamino)propyl,
3-(diethylamino)propyl, 3-(dipropylamino)propyl,
3-(piperidin-1-yl)propyl, 3-(morpholin-1-yl)propyl,
3-(tetrahydropyrrol-1-yl)propyl, 4-(dimethylamino)butyl,
4-(diethylamino)butyl, 4-(dipropylamino)butyl,
4-(piperidin-1-yl)butyl, 4-(morpholin-1-yl)butyl,
4-(tetrahydropyrrol-1-yl)butyl,
2-(2-(2-(2-(dimethylamino)ethoxy)ethoxy)ethoxy)ethyl,
2-(2-(2-(2-(diethylamino)ethoxy)ethoxy)ethoxy)ethyl,
2-(2-(2-(2-(dipropylamino)ethoxy)ethoxy)ethoxy)ethyl,
2-(2-(2-(2-(piperidin-1-yl)ethoxy)ethoxy)ethoxy)ethyl,
2-(2-(2-(2-(morpholin-1-yl)ethoxy)ethoxy)ethoxy)ethyl,
2-(2-(2-(2-(tetrahydropyrrol-1-yl)ethoxy)ethoxy)ethoxy)ethyl,
2-(2-(2-(dimethylamino)ethoxy)ethoxy)ethyl,
2-(2-(2-(diethylamino)ethoxy)ethoxy)ethyl,
2-(2-(2-(dipropylamino)ethoxy)ethoxy)ethyl,
2-(2-(2-(piperidin-1-yl)ethoxy)ethoxy)ethyl,
2-(2-(2-(morpholin-1-yl)ethoxy)ethoxy)ethyl,
2-(2-(2-(tetrahydropyrrol-1-yl)ethoxy)ethoxy)ethyl, 6-purinyl,
mesyl, benzenesulfonyl, pyrazin-2-yl, pyrimidin-2-yl,
2-hydroxyethyl, 2-(2-hydroxyethoxy)ethyl,
2-(2-(2-(2-(2-(2-(2-(2-(2-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)ethoxy)eth-
oxy)ethoxy)ethoxy)ethoxy)ethoxy)ethoxy)ethoxy)ethyl,2-(2-(2-(2-(2-(2-(2-(2-
-(2-hydroxyethoxy)ethoxy)ethoxy)ethoxy)ethoxy)ethoxy)ethoxy)ethoxy)ethyl,
2-(2-(2-(2-(2-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)ethoxy)ethoxy)ethoxy)e-
thoxy)ethyl,
2-(2-(2-(2-(2-(2-(2-(2-(2-methoxyethoxy)ethoxy)ethoxy)ethoxy)ethoxy)ethox-
y)ethoxy)ethoxy)ethyl, and
2-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)ethyl.
30. The method of claim 23, wherein NR.sup.2R.sup.3 is selected
from the group consisting of piperidin-1-yl, morpholin-1-yl,
tetrahydropyrrol-1-yl, (4-(2-hydroxyethyl))piperazin-1-yl,
(4-methyl)piperazin-1-yl, (4-ethyl)piperazin-1-yl,
(4-propyl)piperazin-1-yl, 4-(3-hydroxypropyl)piperazin-1-yl,
3-(morpholin-1-yl)propyl, adenin-1-yl,
(4-(3-(dimethylamino)propyl)piperazin)-1-yl,
(4-(2-(dimethylamino)ethyl)piperazin)-1-yl,
(4-(3-(diethylamino)propyl)piperazin)-l-yl,
(4-(2-(diethylamino)ethyl)piperazin)-1-yl,
(4-(2-(piperidin-1-yl)ethyl)piperazin)-1-yl,
(4-(3-(piperidin-1-yl)propyl)piperazin)-1-yl,
(4-(2-(morpholin-1-yl)ethyl)piperazin)-1-yl,
(4-(3-(morpholin-1-yl)propyl)piperazin)-1-yl,
(4-(2-(tetrahydropyrrol-1-yl)ethyl)piperazin)-1-yl, and
(4-(3-(tetrahydropyrrol-1-yl)propyl)piperazin)-1-yl.
31. The method of claim 23, wherein the compound is selected from
the group consisting of: ##STR00246## wherein W is O, R.sup.1 is
OCH.sub.3, R.sup.4 is CH.sub.3, R.sup.5 is OH, n and
NR.sup.2R.sup.3 are shown in the following table: TABLE-US-00022 o.
NR.sup.2R.sup.3 ##STR00247## ##STR00248## ##STR00249## ##STR00250##
##STR00251## ##STR00252## ##STR00253## ##STR00254## ##STR00255## 0
##STR00256## 1 ##STR00257## 2 ##STR00258## 3 ##STR00259## 4
##STR00260## 5 ##STR00261## 6 ##STR00262## 7 ##STR00263## 8
##STR00264## 9 ##STR00265## 0 ##STR00266## 1 ##STR00267## 2
##STR00268## 3 ##STR00269## 4 ##STR00270## 5 ##STR00271## 6
##STR00272## 7 ##STR00273## 8 ##STR00274## 9 ##STR00275## 0
##STR00276## 1 ##STR00277## 2 ##STR00278## 3 ##STR00279## 4
##STR00280## 5 ##STR00281## 6 ##STR00282## 7 ##STR00283## 8
##STR00284## 9 ##STR00285## 0 ##STR00286## 1 ##STR00287## 2
##STR00288## 3 ##STR00289## 4 ##STR00290## 5 ##STR00291## 6
##STR00292## 7 ##STR00293## 8 ##STR00294## 9 ##STR00295## 0
##STR00296## 1 ##STR00297## 2 ##STR00298## 3 ##STR00299## 4
##STR00300## 5 ##STR00301## 6 ##STR00302## 7 ##STR00303## 8
##STR00304## 9 ##STR00305## 0 ##STR00306## 1 ##STR00307## 2
##STR00308## 3 ##STR00309## 4 ##STR00310## 5 ##STR00311## 6
NH.sub.2 7 NH.sub.2 8 NHCH.sub.3 9 NHCH.sub.3 0 N(CH.sub.3).sub.2 1
N(CH.sub.3).sub.2 2 ##STR00312## 3 ##STR00313## 4 ##STR00314## 5
##STR00315## 8 ##STR00316## 0 ##STR00317## 1 ##STR00318## 2
##STR00319## 3 ##STR00320## 4 ##STR00321## 5 ##STR00322##
77)
10-((3'-(pyrrol-1-yl)-2',3',6'-trideoxy-alphal-L-lyxo-hexylpyranyl)ox-
y)-7,8,9,10-tetrahydro-6,8,11-trihydroxy-13-oxo-14-(4-((4-(2-hydroxy)ethyl-
)piperazin-1-yl)-4-oxo-butyrato)-1-methoxy-5,12-naphthalenedione
acetate; 78)
10-((3'-(pyrrol-1-yl)-2',3',6'-trideoxy-alphal-L-lyxo-hexylpyranyl)ox-
y)-7,8,9,10-tetrahydro-6,8,11-trihydroxy-13-oxo-14-(4-((3-(morpholin-1-yl)-
propyl)amino)-4-oxo-butyrato)-1-methoxy-5,12-naphthalenedione
acetate; 79)
10-((3'-(pyrrol-1-yl)-2',3',6'-trideoxy-alphal-L-lyxo-hexylpyranyl)oxy)-7-
,8,9,10-tetrahydro-6,8,11-trihydroxy-13-oxo-14-(4-((4-methyl)piperazin-1-y-
l)amino)-4-oxo-butyrato)-1-methoxy-5,12-naphthalenedione acetate;
80)
10-((3'-(pyrrol-1-yl)-2',3',6'-trideoxy-alphal-L-lyxo-hexylpyranyl)oxy)-7-
,8,9,10-tetrahydro-6,8,11-trihydroxy-13-oxo-14-(4-(4-ethylpiperazin-1-yl)--
4-oxo-butyrato)-1-methoxy-5,12-naphthalenedione acetate; 81)
10-((3'-(pyrrol-1-yl)-2',3',6'-trideoxy-alphal-L-lyxo-hexylpyranyl)oxy)-7-
,8,9,10-tetrahydro-6,8,11-trihydroxy-13-oxo-14-(4-((3-(4-methylpiperazin-1-
-yl)propyl)amino)-4-oxo-butyrato)-1-methoxy-5,12-naphthalenedione
acetate; 82)
10-((3'-(pyrrol-1-yl)-2',3',6'-trideoxy-alphal-L-lyxo-hexylpyranyl)ox-
y)-7,8,9,10-tetrahydro-6,8,11-trihydroxy-13-oxo-14-(4-((pyridin-4-yl)methy-
l)amino)-4-oxo-butyrato)-1-methoxy-5,12-naphthalenedione acetate;
83)
10-((3'-(pyrrol-1-yl)-2',3',6'-trideoxy-alphal-L-lyxo-hexylpyranyl)oxy)-7-
,8,9,10-tetrahydro-6,8,11-trihydroxy-13-oxo-14-(4-((pyridin-3-yl)methyl)am-
ino)-4-oxo-butyrato)-1-methoxy-5,12-naphthalenedione acetate; 84)
10-((3'-(pyrrol-1-yl)-2',3',6'-trideoxy-alphal-L-lyxo-hexylpyranyl)oxy)-7-
,8,9,10-tetrahydro-6,8,11-trihydroxy-13-oxo-14-(4-(2-(pyridin-2-yl)ethyl)a-
mino)-4-oxo-butyrato)-1-methoxy-5,12-naphthalenedione acetate; 85)
10-((3'-(pyrrol-1-yl)-2',3',6'-trideoxy-alphal-L-lyxo-hexylpyranyl)oxy)-7-
,8,9,10-tetrahydro-6,8,11-trihydroxy-13-oxo-14-(4-(2-(pyridin-3-yl)ethyl)a-
mino)-4-oxo-butyrato)-1-methoxy-5,12-naphthalenedione acetate; 86)
10-((3'-(pyrrol-1-yl)-2',3',6'-trideoxy-alphal-L-lyxo-hexylpyranyl)oxy)-7-
,8,9,10-tetrahydro-6,8,11-trihydroxy-13-oxo-14-(4-(2-(pyridin-4-yl)ethyl)a-
mino)-4-oxo-butyrato)-1-methoxy-5,12-naphthalenedione acetate; 87)
10-((3'-(pyrrol-1-yl)-2',3',6'-trideoxy-alphal-L-lyxo-hexylpyranyl)oxy)-7-
,8,9,10-tetrahydro-6,8,11-trihydroxy-13-oxo-14-(4-(4-(3-(dimethylamino)pro-
pyl)piperazin-1-yl)-4-oxo-butyrato)-1-methoxy-5,12-naphthalenedione
acetate; 89)
10-((3'-(pyrrol-1-yl)-2',3',6'-trideoxy-alphal-L-lyxo-hexylpyranyl)oxy)-7-
,8,9,10-tetrahydro-6,8,11-trihydroxy-13-oxo-14-(4-((2-(morpholin-1-yl)ethy-
l)amino)-4-oxo-butyrato)-1-methoxy-5,12-naphthalenedione acetate;
96)
10-((3'-(pyrrol-1-yl)-2',3',6'-trideoxy-alphal-L-lyxo-hexylpyranyl)oxy)-7-
,8,9,10-tetrahydro-6,8,11-trihydroxy-13-oxo-14-(4-((2-(morpholin-1-yl)ethy-
l)amino)-4-oxo-butyrato)-1-methoxy-5,12-naphthalenedione phosphate;
and 99)
10-((3'-(pyrrol-1-yl)-2',3',6'-trideoxy-alphal-L-lyxo-hexylpyranyl)ox-
y)-7,8,9,10-tetrahydro-6,8,11-trihydroxy-13-oxo-14-(4-((2-(morpholin-1-yl)-
ethyl)amino)-4-oxo-butyrato)-1-methoxy-5,12-naphthalenedione
hydrochloride.
32. The method of claim 23, wherein the formulation is a
formulation for injection.
Description
FIELD
[0001] The present application relates to the field of organic
compounds and medicinal chemistry. In particular, the present
application relates to tetracyclic anthraquinone derivatives,
preparation processes and applications thereof.
BACKGROUND
[0002] Tetracyclic anthraquinone antibiotics, in particular
doxorubicin and daunorubicin, are widely used anticancer drugs.
Doxorubicin has significant curative effects on lots of solid
tumors including liver cancer, gastric cancer, breast cancer, lung
cancer, ovary cancer and various leukemias. Daunorubicin is one of
the most effective drugs for treating leukemia. However, due to
their side effects such as severe myelosuppression, cardiac
toxicity, adverse responses in digestive tracts and the like, their
clinical applications are somewhat restricted. Up to now, lots of
derivatives of tetracyclic anthraquinones have already been
separated from the nature or artificially synthesized.
SUMMARY
[0003] One aspect of the present application relates to a compound
represented by formula (I) and a pharmaceutically acceptable salt
thereof,
##STR00002##
[0004] wherein:
[0005] R.sup.1 is selected from the group consisting of H,
optionally substituted alkyl, and optionally substituted
alkoxy;
[0006] R.sup.2 is selected from the group consisting of H,
optionally substituted aryl, optionally substituted heteroaryl,
optionally substituted (alkyleneoxy).sub.malkyl, optionally
substituted heterocyclyl, optionally substituted alkyl, and
optionally substituted sulfonyl;
[0007] R.sup.3 is selected from the group consisting of H,
optionally substituted aryl, optionally substituted heteroaryl,
optionally substituted (alkyleneoxy).sub.malkyl, optionally
substituted heterocyclyl, optionally substituted alkyl, and
optionally substituted sulfonyl;
[0008] or NR.sup.2R.sup.3 represents optionally substituted
heterocyclyl;
[0009] m is selected from the group consisting of 0, 1, 2, 3, 4, 5,
6, 7, 8, 9, 10, and 11;
[0010] W is selected from the group consisting of O and NH;
[0011] R.sup.4 is selected from the group consisting of H, F, and
optionally substituted alkyl;
[0012] R.sup.5 is selected from the group consisting of H, F,
optionally substituted alkyl and OR.sup.6, wherein R.sup.6 is
selected from the group consisting of H and
tetrahydropyran-2-yl;
[0013] n is selected from the group consisting of 1, 2 and 3.
[0014] Another aspect of the present application relates to a
compound represented by formula (I) and a pharmaceutically
acceptable salt thereof,
##STR00003##
[0015] wherein:
[0016] R.sup.1 is selected from the group consisting of H,
C.sub.1-4alkyl, and C.sub.1-4alkoxy;
[0017] R.sup.2 is selected from the group consisting of H, aryl,
heteroaryl, (C.sub.1-4alkyleneoxy).sub.mC.sub.1-4alkyl,
heterocyclyl, C.sub.1-4alkyl, and sulfonyl;
[0018] R.sup.3 is selected from the group consisting of H, aryl,
heteroaryl, (C.sub.1-4alkyleneoxy).sub.mC.sub.1-4alkyl,
heterocyclyl, C.sub.1-4alkyl, and sulfonyl;
[0019] or NR.sup.2R.sup.3 represents heterocyclyl;
[0020] m is selected from the group consisting of 0, 1, 2, 3, 4, 5,
6, 7, 8, 9, 10, and 11;
[0021] W is selected from the group consisting of 0 and NH;
[0022] R.sup.4 is selected from the group consisting of H, F and
C.sub.1-4alkyl;
[0023] R.sup.5 is selected from the group consisting of H, F,
C.sub.1-4alkyl and OR.sup.6, wherein R.sup.6 is selected from the
group consisting of H and tetrahydropyran-2-yl; and
[0024] n is selected from the group consisting of 1, 2 and 3.
[0025] Yet another aspect of the present application relates to a
compound selected from the group consisting of:
TABLE-US-00001 ##STR00004## No. n NR.sup.2R.sup.3 1 3 ##STR00005##
2 2 ##STR00006## 3 3 ##STR00007## 4 2 ##STR00008## 5 3 ##STR00009##
6 2 ##STR00010## 7 3 ##STR00011## 8 2 ##STR00012## 9 3 ##STR00013##
10 2 ##STR00014## 11 3 ##STR00015## 12 2 ##STR00016## 13 3
##STR00017## 14 2 ##STR00018## 15 3 ##STR00019## 16 2 ##STR00020##
17 3 ##STR00021## 18 2 ##STR00022## 19 3 ##STR00023## 20 2
##STR00024## 21 3 ##STR00025## 22 2 ##STR00026## 23 3 ##STR00027##
24 2 ##STR00028## 25 3 ##STR00029## 26 2 ##STR00030## 27 3
##STR00031## 28 2 ##STR00032## 29 3 ##STR00033## 30 2 ##STR00034##
31 3 ##STR00035## 32 2 ##STR00036## 33 3 ##STR00037## 34 2
##STR00038## 35 3 ##STR00039## 36 2 ##STR00040## 37 3 ##STR00041##
38 2 ##STR00042## 39 3 ##STR00043## 40 2 ##STR00044## 41 3
##STR00045## 42 2 ##STR00046## 43 3 ##STR00047## 44 2 ##STR00048##
45 3 ##STR00049## 46 2 ##STR00050## 47 3 ##STR00051## 48 2
##STR00052## 49 3 ##STR00053## 50 2 ##STR00054## 51 3 ##STR00055##
52 2 ##STR00056## 53 3 ##STR00057## 54 2 ##STR00058## 55 3
##STR00059## 56 2 ##STR00060## 57 3 ##STR00061## 58 2 ##STR00062##
59 3 ##STR00063## 60 2 ##STR00064## 61 3 ##STR00065## 62 2
##STR00066## 63 3 ##STR00067## 64 2 ##STR00068## 65 3 ##STR00069##
66 2 NH.sub.2 67 3 NH.sub.2 68 2 NHCH.sub.3 69 3 NHCH.sub.3 70 2
N(CH.sub.3).sub.2 71 3 N(CH.sub.3).sub.2 72 2 ##STR00070## 73 3
##STR00071## 74 2 ##STR00072## 75 2 ##STR00073## 88 2 ##STR00074##
90 2 ##STR00075## 91 2 ##STR00076## 92 2 ##STR00077## 93 2
##STR00078## 94 2 ##STR00079## 95 2 ##STR00080##
[0026]
10-((3'-(pyrrol-1-yl)-2',3',6'-trideoxy-alphal-L-lyxo-hexylpyranyl-
)oxy)-7,8,9,10-tetrahydro-6,8,11-trihydroxy-13-oxo-14-(4-((4-(2-hydroxy)et-
hyl)piperazin-1-yl)-4-oxo-butyrato)-1-methoxy-5,12-naphthalenedione
acetate; [0027] 78)
10-((3'-(pyrrol-1-yl)-2',3',6'-trideoxy-alphal-L-lyxo-hexylpyranyl)oxy)-7-
,8,9,10-tetrahydro-6,8,11-trihydroxy-13-oxo-14-(4-((3-(morpholin-1-yl)prop-
yl)amino)-4-oxo-butyrato)-1-methoxy-5,12-naphthalenedione acetate;
[0028] 79)
10-((3'-(pyrrol-1-yl)-2',3',6'-trideoxy-alphal-L-lyxo-hexylpyranyl)ox-
y)-7,8,9,10-tetrahydro-6,8,11-trihydroxy-13-oxo-14-(4-((4-methyl)piperazin-
-1-yl)amino)-4-oxo-butyrato)-1-methoxy-5,12-naphthalenedione
acetate; [0029] 80)
10-((3'-(pyrrol-1-yl)-2',3',6'-trideoxy-alphal-L-lyxo-hexylpyranyl)oxy)-7-
,8,9,10-tetrahydro-6,8,11-trihydroxy-13-oxo-14-(4-(4-ethylpiperazin-1-yl)--
4-oxo-butyrato)-1-methoxy-5,12-naphthalenedione acetate; [0030] 81)
10-((3'-(pyrrol-1-yl)-2',3',6'-trideoxy-alphal-L-lyxo-hexylpyranyl)oxy)-7-
,8,9,10-tetrahydro-6,8,11-trihydroxy-13-oxo-14-(4-((3-(4-methylpiperazin-1-
-yl)propyl)amino)-4-oxo-butyrato)-1-methoxy-5,12-naphthalenedione
acetate; [0031] 82)
10-((3'-(pyrrol-1-yl)-2',3',6'-trideoxy-alphal-L-lyxo-hexylpyranyl)oxy)-7-
,8,9,10-tetrahydro-6,8,11-trihydroxy-13-oxo-14-(4-((pyridin-4-yl)methyl)am-
ino)-4-oxo-butyrato)-1-methoxy-5,12-naphthalenedione acetate;
[0032] 83)
10-((3'-(pyrrol-1-yl)-2',3',6'-trideoxy-alphal-L-lyxo-hexylpyranyl)oxy)-7-
,8,9,10-tetrahydro-6,8,11-trihydroxy-13-oxo-14-(4-((pyridin-3-yl)methyl)am-
ino)-4-oxo-butyrato)-1-methoxy-5,12-naphthalenedione acetate;
[0033] 84)
10-((3'-(pyrrol-1-yl)-2',3',6'-trideoxy-alphal-L-lyxo-hexylpyranyl)oxy)-7-
,8,9,10-tetrahydro-6,8,11-trihydroxy-13-oxo-14-(4-(2-(pyridin-2-yl)ethyl)a-
mino)-4-oxo-butyrato)-1-methoxy-5,12-naphthalenedione acetate;
[0034] 85)
10-((3'-(pyrrol-1-yl)-2',3',6'-trideoxy-alphal-L-lyxo-hexylpyranyl)oxy)-7-
,8,9,10-tetrahydro-6,8,11-trihydroxy-13-oxo-14-(4-(2-(pyridin-3-yl)ethyl)a-
mino)-4-oxo-butyrato)-1-methoxy-5,12-naphthalenedione acetate;
[0035] 86)
10-((3'-(pyrrol-1-yl)-2',3',6'-trideoxy-alphal-L-lyxo-hexylpyranyl)oxy)-7-
,8,9,10-tetrahydro-6,8,11-trihydroxy-13-oxo-14-(4-(2-(pyridin-4-yl)ethyl)a-
mino)-4-oxo-butyrato)-1-methoxy-5,12-naphthalenedione acetate;
[0036] 87)
10-((3'-(pyrrol-1-yl)-2',3',6'-trideoxy-alphal-L-lyxo-hexylpyranyl)oxy)-7-
,8,9,10-tetrahydro-6,8,11-trihydroxy-13-oxo-14-(4-(4-(3-(dimethylamino)pro-
pyl)piperazin-1-yl)-4-oxo-butyrato)-1-methoxy-5,12-naphthalenedione
acetate; [0037] 89)
10-((3'-(pyrrol-1-yl)-2',3',6'-trideoxy-alphal-L-lyxo-hexylpyranyl)oxy)-7-
,8,9,10-tetrahydro-6,8,11-trihydroxy-13-oxo-14-(4-((2-(morpholin-1-yl)ethy-
l)amino)-4-oxo-butyrato)-1-methoxy-5,12-naphthalenedione acetate;
[0038] 96)
10-((3'-(pyrrol-1-yl)-2',3',6'-trideoxy-alphal-L-lyxo-hexylpyranyl)ox-
y)-7,8,9,10-tetrahydro-6,8,11-trihydroxy-13-oxo-14-(4-((2-(morpholin-1-yl)-
ethyl)amino)-4-oxo-butyrato)-1-methoxy-5,12-naphthalenedione
phosphate; and [0039] 99)
10-((3'-(pyrrol-1-yl)-2',3',6'-trideoxy-alphal-L-lyxo-hexylpyranyl)oxy)-7-
,8,9,10-tetrahydro-6,8,11-trihydroxy-13-oxo-14-(4-((2-(morpholin-1-yl)ethy-
l)amino)-4-oxo-butyrato)-1-methoxy-5,12-naphthalenedione
hydrochloride.
[0040] Still another aspect of the present application relates to a
process for preparing a compound represented by formula (I),
comprising:
##STR00081##
[0041] reacting a compound represented by formula (II) with a
compound represented by formula (III) in the presence of a
condensation agent to obtain the compound represented by formula
(I),
[0042] wherein:
[0043] in the compound represented by formula (I), R.sup.1 is
selected from the group consisting of H, C.sub.1-4alkyl, and
C.sub.1-4alkoxy; R.sup.2 is selected from the group consisting of
H, aryl, heteroaryl, (C.sub.1-4alkyleneoxy).sub.mC.sub.1-4alkyl,
heterocyclyl, C.sub.1-4alkyl, and sulfonyl; R.sup.3 is selected
from the group consisting of H, aryl, heteroaryl,
(C.sub.1-4alkyleneoxy).sub.mC.sub.1-4alkyl, heterocyclyl,
C.sub.1-4alkyl, and sulfonyl; or NR.sup.2R.sup.3 represents
heterocyclyl; m is selected from the group consisting of 0, 1, 2,
3, 4, 5, 6, 7, 8, 9, 10, and 11; W is selected from the group
consisting of 0 and NH; R.sup.4 is selected from the group
consisting of H, F and C.sub.1-4alkyl; R.sup.5 is selected from the
group consisting of H, F, C.sub.1-4alkyl and OR.sup.6, wherein
R.sup.6 is selected from the group consisting of H and
tetrahydropyran-2-yl; and n is selected from the group consisting
of 1, 2 and 3;
[0044] groups represented by R.sup.1, W, R.sup.4, R.sup.5 in the
compound represented by formula (II) are the same as groups
represented by R.sup.1, W, R.sup.4, R.sup.5 in the compound
represented by formula (I);
[0045] n in the compound represented by formula (III) has the same
meanings as n in the compound represented by formula (I); groups
represented by R.sup.7 and R.sup.8 in the compound represented by
formula (III) are the same as groups represented by R.sup.2 and
R.sup.3 in the compound represented by formula (I), provided that
groups represented by R.sup.7 and R.sup.8 do not comprise NH or
NH.sub.2; when groups represented by R.sup.7 and R.sup.8 comprise
NH or NH.sub.2, the compound represented by formula (III) has an
amino-protecting group at N-terminus, and is subject to a
deprotection reaction to obtain the compound represented by formula
(I).
[0046] Yet another aspect of the present application relates to a
pharmaceutical composition comprising a compound represented by
formula (I) or a pharmaceutically acceptable salt thereof, and a
pharmaceutically acceptable carrier.
[0047] Still another aspect of the present application relates to a
formulation comprising a compound represented by formula (I) or a
pharmaceutically acceptable salt thereof, and a pharmaceutically
acceptable carrier.
[0048] Yet another aspect of the present application relates to a
method for treating and/or preventing tumor and/or cancer,
comprising administering to a subject in need thereof a
therapeutically effective amount of a compound represented by
formula (I) or a pharmaceutically acceptable salt thereof, or
administering a therapeutically effective amount of a
pharmaceutical composition comprising a compound represented by
formula (I) or a pharmaceutically acceptable salt and a
pharmaceutically acceptable carrier, or administering a
therapeutically effective amount of a formulation comprising a
compound represented by formula (I) or a pharmaceutically
acceptable salt and a pharmaceutically acceptable carrier.
[0049] The compound and a salt thereof according to the present
application possess good anticancer and/or antitumor activities,
and good water solubility and stability, as well as good tolerance
in animal bodies. Therefore, they are prone to being developed as
clinical drugs.
BRIEF DESCRIPTION OF DRAWINGS
[0050] FIG. 1 shows effects of 3'-pyrrolyldoxorubicin on the body
weight of experimental animals.
[0051] FIG. 2 shows effects of the compound in Example 96 according
to the present application on the body weight of experimental
animals.
[0052] FIG. 3 shows effects of
3'-pyrrolyldoxorubicin-14-oxo-succinic acid monoester on the body
weight of experimental animals.
DETAILED DESCRIPTION
[0053] In the following description, certain specific details are
included to provide a thorough understanding of various disclosed
embodiments. One of ordinary skill in the relevant art, however,
will recognize that the embodiments may be practiced without one or
more of these specific details, or with other methods, components,
materials etc.
[0054] Unless the context required otherwise, throughout the
specification and claims which follows, the term "comprise" and
variations thereof, such as "comprises" and "comprising" are to be
construed in an open, inclusive sense, which is as "include, but
not limited to".
[0055] Reference throughout this specification to "one embodiment",
or "an embodiment", or "in another embodiment", or "in some
embodiments" means that a particular referent feature, structure or
characteristics described in connection with the embodiment is
included in at least one embodiment. Therefore, the appearance of
the phrases "in one embodiment" or "in the embodiment" or "in
another embodiment" or "in some embodiments" in various places
throughout this specification are not necessarily all referring to
the same embodiment. Moreover, the particular features, structures
or characteristics may be combined in any suitable manner in one or
more embodiments.
[0056] It should be noted that, as used in this specification and
the appended claims, the singular forms "a", "an" and "the" include
plural referents unless the context clearly stated otherwise.
Therefore, for example, a reaction comprising "a catalyst"
comprises one catalyst, two or more catalysts. It should be also
noted that the use of "or" means "and/or" unless stated
otherwise.
Definitions
[0057] Certain chemical groups named herein are preceded by a
shorthand notation indicating the total number of carbon atoms that
are to be found in the indicated chemical group. For example,
C.sub.7-C.sub.12alkyl describes an alkyl group, as defined below,
having a total of 7 to 12 carbon atoms, and C.sub.4-C.sub.12
cyclohydrocarbylalkyl describes a cyclohydrocarbylalkyl group, as
defined below, having a total of 4 to 12 carbon atoms. The total
number of carbon atoms in the shorthand notation does not include
the carbons that may exist in the substituents of the groups
described.
[0058] Accordingly, as used in the specification and appended
claims, unless specified to the contrary, the following terms have
the meanings indicated:
[0059] The term "alkyl", as used herein, refers to a straight or
branched hydrocarbon chain group consisting solely of carbon and
hydrogen, containing no unsaturated bond, having from one to twelve
carbon atoms, preferably one to eight or one to six carbon atoms,
and which is attached to the rest of the molecule by a single bond,
e.g., methyl, ethyl, n-propyl, 1-methylethyl (iso-propyl), n-butyl,
n-pentyl, 1,1-dimethylethyl (tert-butyl), 3-methylhexyl,
2-methylhexyl, and the like.
[0060] Alkyl group may have one to twelve carbon atoms (whenever it
appears in the present application, a numerical range such as "one
to twelve" refers to each integer in the given number range; e.g.
"one to twelve carbon atoms" means that the alkyl group may consist
of one carbon atom, two carbon atoms, three carbon atoms, etc., up
to and including twelve carbon atoms, although the present
definition also covers the occurrence of term "alkyl" where no
numerical range is designated). Alkyl group may also be a medium
sized alkyl having one to ten carbon atoms. Alkyl group may also be
a lower alkyl having one to five carbon atoms. Alkyl group of
compounds of the present application may be designated as
"C.sub.1-4 alkyl" or similar designations. By way of example only,
"C.sub.1-4 alkyl" indicates that there are one to four carbon atoms
in the alkyl chain, i.e. the alkyl chain is selected from the group
consisting of methyl, ethyl, propyl, isopropyl, n-butyl, iso-butyl,
sec-butyl, or t-butyl.
[0061] Alkyl group may be optionally substituted, i.e. substituted
or unsubstituted. When substituted, the substituted group(s)
is(are) individually and independently selected from the group
consisting of cyclohydrocarbyl, aryl, heteroaryl, heteroalicyclyl,
hydroxy, alkoxy, aryloxy, mercapto, allkylthio, arylthio, cyano,
halo, carbonyl, thiocarbonyl, O-carbamyl, N-carbamyl,
O-thiocarbamyl, N-thiocarbamyl, C-amido, N-amido, S-sulfinamido,
N-sulfinamido, C-carboxyl, O-carboxyl, isocyanato, thiocyano,
isothiocyanato, nitro, silyl, trihalomethanesulfonyl, --NR'R'' or
amino including mono- and bi-substituted amino group, and the
protected derivatives thereof. Typical hydrocarbyl groups include,
but are not limited to, methyl, ethyl, propyl, isopropyl, butyl,
isobutyl, t-butyl, pentyl, hexyl, ethenyl, propenyl, buenyl,
cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.
Whenever a substituent is described as being "optionally
substituted", that substituent may be substituted with one of the
above substituents.
[0062] "C.sub.1-4 alkyl" refers to an alkyl group as defined above
containing one to four carbon atoms. C.sub.1-4 alkyl group may be
optionally substituted as defined for alkyl group.
[0063] "C.sub.1-6 alkyl" refers to an alkyl group as defined above
containing one to six carbon atoms. C.sub.1-6 alkyl group may be
optionally substituted as defined for alkyl group.
[0064] "C.sub.1-12 alkyl" refers to an alkyl group as defined above
containing one to twelve carbon atoms. C.sub.1-12 alkyl group may
be optionally substituted as defined for alkyl group.
[0065] "C.sub.2-6 alkyl" refers to an alkyl group as defined above
containing two to six carbon atoms. C.sub.2-6 alkyl group may be
optionally substituted as defined for alkyl group.
[0066] "C.sub.3-6 alkyl" refers to an alkyl as defined above
containing three to six carbon atoms. C.sub.3-6 alkyl group may be
optionally substituted as defined for alkyl group.
[0067] "C.sub.3-12 alkyl" refers to an alkyl as defined above
containing three to twelve carbon atoms. C.sub.3-12 alkyl group may
be optionally substituted as defined for alkyl group.
[0068] "C.sub.6-12 alkyl" refers to an alkyl as defined above
containing six to twelve carbon atoms. C.sub.6-12 alkyl group may
be optionally substituted as defined for alkyl group.
[0069] "C.sub.7-12 alkyl" refers to an alkyl as defined above
containing seven to twelve carbon atoms. C.sub.7-12 alkyl group may
be optionally substituted as defined for alkyl group.
[0070] In some embodiments, the alkyl group is C.sub.1-12
alkyl.
[0071] In some embodiments, the alkyl group is C.sub.1-8 alkyl.
[0072] In some embodiments, the alkyl group is C.sub.1-6 alkyl.
[0073] In some embodiments, the alkyl group is C.sub.1-4 alkyl.
[0074] "Alkoxy", as used herein, refers to the formula --OR,
wherein R is an alkyl group defined as above, e.g. methoxy, ethoxy,
n-propoxy, 1-methyl ethoxy (isopropoxy), n-butoxy, isobutoxy,
sec-butoxy, t-butoxy, amoxy, t-amoxy, and the like.
[0075] In some embodiments, the alkoxy group is C.sub.1-12
alkoxy.
[0076] In some embodiments, the alkoxy group is C.sub.1-8
alkoxy.
[0077] In some embodiments, the alkoxy group is C.sub.1-6
alkoxy.
[0078] In some embodiments, the alkoxy group is C.sub.1-4
alkoxy.
[0079] "Alkylene", as used herein, refers to a straight or branched
divalent hydrocarbon chain group consisting solely of carbon and
hydrogen and having from one to eight carbon atoms, which is linked
with the other moiety of the molecule and a residual group, e.g.
methylene, ethylene, propylene, n-butylene. Alkylene chain can be
linked with the other moiety of the molecule and the residual group
via one carbon atom in the chain or any two carbon atoms in the
chain.
[0080] In some embodiments, the alkylene group is C.sub.1-12
alkylene.
[0081] In some embodiments, the alkylene group is C.sub.1-8
alkylene.
[0082] In some embodiments, the alkylene group is C.sub.1-6
alkylene.
[0083] In some embodiments, the alkylene group is C.sub.1-4
alkylene.
[0084] "Alkyleneoxy", as used herein, refers to the formula --OR,
wherein R is an alkylene group defined as above, e.g. methyleneoxy,
ethyleneoxy, n-propyleneoxy, isopropyleneoxy, n-butyleneoxy,
isobutyleneoxy, sec-butyleneoxy, t-butyleneoxy, amyleneoxy,
t-amyleneoxy, and the like.
[0085] In some embodiments, the alkyleneoxy group is
O(C.sub.1-12)alkylene.
[0086] In some embodiments, the alkyleneoxy group is
O(C.sub.1-8)alkylene.
[0087] In some embodiments, the alkyleneoxy group is
O(C.sub.1-6)alkylene.
[0088] In some embodiments, the alkyleneoxy group is
O(C.sub.1-4)alkylene.
[0089] "Aryl", as used herein, refers to a carbocycle (full carbon)
or two or more fused rings (rings sharing with two adjacent carbon
atoms), having completely delocalized .pi. electron system.
Examples of aryl group include, but are not limited to, fluorenyl,
phenyl and naphthyl. The aryl group may have, for example, five to
twelve carbon atoms. The aryl group of the present application may
be substituted or unsubstituted. Where substituted, hydrogen
atom(s) is(are) substituted with one or more substituents
independently selected from the group consisting of alkyl,
cyclohydrocarbyl, aryl, heteroaryl, heteroalicyclyl, hydroxy,
protected hydroxy, alkoxy, aryloxy, mercapto, alkylthio, arylthio,
cyano, halo, carbonyl, thiocarbonyl, O-carbamyl, N-carbamyl,
O-thiocarbamyl, N-thiocarbamyl, C-amido, N-amido, S-sulfinamido,
N-sulfinamido, C-carboxyl, protected C-carboxyl, O-carboxyl,
isocyanato, thiocyano, isothiocyanato, nitro, silyl,
trihalomethanesulfonyl, --NR'R'' (R' and R'' are alkyl groups
defined herein) and protected amino.
[0090] In some embodiments, the aryl group is C.sub.6-C.sub.18
aryl.
[0091] In some embodiments, the aryl group is C.sub.6-C.sub.12
aryl.
[0092] In some embodiments, the aryl group is C.sub.6-C.sub.10
aryl.
[0093] "Heteroaryl (aromatic heterocyclyl)" refers to a five- to
eighteen-membered aromatic ring group, containing one to seventeen
carbon atoms and one to ten heteroatoms selected from the group
consisting of nitrogen, oxygen and sulphur. For the purpose of the
present invention, the heteroaryl may be monocyclic, bicyclic,
tricyclic or tetracyclic ring system, which may comprise fused or
bridged ring system. Moreover, nitrogen, carbon or sulphur atom in
the heteroaryl group may be optionally oxidized, and the nitrogen
atom may be optionally quaternized. The examples include, but are
not limited to, azepinyl, acridinyl, benzimidazolyl,
benzothiazolyl, benzoindolyl, benzodioxolanyl, benzofuranyl,
benzoxazolyl, benzothiazolyl, benzothiadiazolyl,
benzo[b][1,4]dioxepanyl, 1,4-benzodioxanyl, benzonaphthofuranyl,
benzodioxolanyl, benzodioxadienyl, benzopyranyl, benzopyronyl,
benzofuranyl, benzofuranonyl, benzothienyl, benzotriazolyl,
benzo[4,6]imidazo[1,2-a]pyridyl, carbazolyl, cinnolinyl,
dibenzofuranyl, dibenzothienyl, furanyl, furanonyl, isothiazolyl,
imidazolyl, indazolyl, indolyl, indazolyl, isoindolyl, indolinyl,
isoindolinyl, isoquinolyl, indolizinyl, isoxazolyl, naphthyl,
naphthyridinyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl, oxiranyl,
1-phenyl-1H-pyrrolyl, phenazinyl, phenothiazinyl, phenoxazinyl,
2,3-naphthyridinyl, pteridinyl, purinyl, pyrrolyl, pyrazolyl,
pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrrolyl,
quinazolinyl, quinoxalinyl, quinolyl, quinuclidinyl, isoquinolyl,
tetrahydroquinolyl, thiazolyl, thiadiazolyl, triazolyl, tetrazolyl,
triazinyl and thiophenyl. Unless stated otherwise specifically in
the specification, the term "heteroaryl" is meant to include the
heteroaryl groups which may be optionally substituted with one or
more substituents independently selected from the group consisting
of alkyl, cyclohydrocarbyl, aryl, heteroaryl, heteroalicyclyl,
hydroxy, protected hydroxy, alkoxy, aryloxy, mercapto, alkylthio,
arylthio, cyano, halo, carbonyl, thiocarbonyl, O-carbamyl,
N-carbamyl, O-thiocarbamyl, N-thiocarbamyl, C-amido, N-amido,
S-sulfinamido, N-sulfinamido, C-carboxyl, protected C-carboxyl,
O-carboxyl, isocyanato, thiocyano, isothiocyanato, nitro, silyl,
trihalomethanesulfonyl, --NR'R'' (R' and R'' are alkyl groups
defined herein) and protected amino.
[0094] In some embodiments, the heteroaryl group is C.sub.5-18
heteroaryl.
[0095] In some embodiments, the heteroaryl group is C.sub.5-12
heteroaryl.
[0096] In some embodiments, the heteroaryl group is C.sub.5-10
heteroaryl.
[0097] The term "heterocyclyl", as used herein, refers to a stable
three- to twelve-membered non-aromatic ring group which consists of
carbon atoms and from one to five heteroatoms selected from the
group consisting of nitrogen, oxygen and sulphur. Examples of such
heteroyclyl groups include, but are not limited to, dioxolanyl,
decahydroisoquinolyl, imidazolinyl, imidazolidinyl,
isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl,
octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl,
2-oxopyrrolidinyl, oxazolidinyl, piperidyl, piperazinyl,
4-piperidonyl, pyrrolidinyl, pyrazolidinyl, thiazolidinyl,
tetrahydrofuryl, trithianyl, tetrahydropyranyl, thiomorpholinyl,
thiamorpholinyl, 1-oxo-thiomorpholinyl and
1,1-dioxo-thiomorpholinyl. Unless stated otherwise specifically in
the specification, the term "heterocyclyl" is meant to include the
heterocyclyl groups as defined above, which may be optionally
substituted with one or more substituents selected from the group
consisting of cyclohydrocarbyl, aryl, heteroaryl, heteroalicyclyl,
hydroxy, alkoxy, aryloxy, mercapto, alkylthio, arylthio, cyano,
halo, carbonyl, thiocarbonyl, O-carbamyl, N-carbamyl,
O-thiocarbamyl, N-thiocarbamyl, C-amido, N-amido, S-sulfinamido,
N-sulfinamido, C-carboxyl, O-carboxyl, isocyanato, thiocyanato,
isothiocyanato, nitro, silyl, trihalomethanesulfonyl, --NR'R'' (R'
and R'' are alkyl groups as defined in the present application) or
amino including mono- and di-substituted amino group, and the
protected derivatives thereof.
[0098] In some embodiments, the heterocyclyl group is C.sub.3-18
heterocyclyl.
[0099] In some embodiments, the heterocyclyl group is C.sub.3-12
heterocyclyl.
[0100] In some embodiments, the heterocyclyl group is C.sub.3-10
heterocyclyl.
[0101] "Sulfonyl" refers to --S(.dbd.O).sub.2R group, in which R
may be alkyl, cyclohydrocarbyl, heterocyclyl, aryl, heteroaryl,
etc, as defined above. The examples of sulfonyl groups include, but
are not limited to, --S(.dbd.O).sub.2CH.sub.3 (mesyl),
--S(.dbd.O).sub.2CF.sub.3, --S(.dbd.O).sub.2CH.sub.2CH.sub.3 and
4-methylbenzenesulfonyl (tosyl).
[0102] "Optional" or "optionally" means that the subsequently
described circumstances may or may not occur, and that the
specification includes instances where said event or circumstance
occurs and instances in which it does not. For example, "optionally
substituted aryl" means that the aryl group may or may not be
substituted and that the specification includes the substituted
aryl group and the aryl group which is not substituted.
[0103] "Pharmaceutically acceptable carriers" include without
limitation to any adjuvant, carrier, excipient, glidant, sweetening
agent, diluent, preservative, dye/colorant, flavor enhancer,
surfactant, wetting agent, dispersing agent, suspending agent,
stabilizer, isosmotic agent, solvent, or emulsifier, etc, which
have been approved by the United States Food and Drug
Administration as being acceptable for use in humans or animals and
have no side effects for constituting a pharmaceutical
composition.
[0104] "Pharmaceutically acceptable salts" include both
"pharmaceutically acceptable acid addition salts" and
"pharmaceutically acceptable base addition salts".
[0105] "Pharmaceutically acceptable acid addition salt" refers to
those salts which retain the biological effectiveness and
properties of free bases, which are biologically or otherwise
desirable, and which are formed with inorganic acids such as, but
not limited to hydrochloric acid, hydrobromic acid, sulfuric acid,
nitric acid, phosphoric acid and the like; and organic acids such
as, but not limited to, acetic acid, 2,2-dichloroacetic acid,
adipic acid, alginic acid, ascorbic acid, aspartic acid,
benzenesulfonic acid, benzoic acid, 4-acetamidobenzoic acid,
camphanic acid, camphor-10-sulfonic acid, capric acid, caproic
acid, caprylic acid, carbonic acid, cinnamic acid, citric acid,
cyclamic acid, dodecylsulfuric acid, ethane-1,2-disulfonic acid,
ethanesulfonic acid, 2-hydroxyethanesulfonic acid, formic acid,
fumaric acid, galactaric acid, gentisic acid, glucoheptonic acid,
gluconic acid, glucuronic acid, glutamic acid, glutaric acid,
2-oxo-glutaric acid, glycerophosphoric acid, glycolic acid,
hippuric acid, isobutyric acid, lactic acid, lactobionic acid,
lauric acid, maleic acid, malic acid, malonic acid, mandelic acid,
methanesulfonic acid, mucic acid, naphthalene-1,5-disulfonic acid,
naphthalene-2-sulfonic acid, 1-hydroxy-2-naphthoic acid, nicotinic
acid, oleinic acid, orotic acid, oxalic acid, palmitic acid, pamoic
acid, propionic acid, pyroglutamic acid, pyruvic acid, salicylic
acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic
acid, tartaric acid, thiocyanic acid, p-toluenesulfonic acid,
trifluoroacetic acid, undecylenic acid and the like.
[0106] "Pharmaceutically acceptable base addition salt" refers to
those salts which retain the biological effectiveness and
properties of the free acids, which are biologically or otherwise
desirable. These salts are prepared from addition of an inorganic
base or an organic base into the free acid. Salts derived from
inorganic bases include, but are not limited to, sodium, potassium,
lithium, ammonium, calcium, magnesium, iron, zinc, copper,
manganese, aluminum slats, and the like. Preferred inorganic salts
are the ammonium, sodium, potassium, calcium, and magnesium salts.
Salts derived from organic bases include, but are not limited to,
slats of primary, secondary and tertiary amines, substituted amines
including naturally occurring substituted amines, cyclic amines and
basic ion exchange resins, such as ammonia, isopropylamine,
trimethylamine, diethylamine, triethylamine, tripropylamine,
diethanolamine, ethanolamine, deanol, 2-dimethylaminoethanol,
2-diethylaminethanol, dicyclohexylamine, lysine, arginine,
histidine, caffeine, procaine, hydrabamine, choline, betaine,
benzylamine, phenylethylenediamine, ethylenediamine, glucosamine,
methylglucosamine, theobromine, triethanolamine, trometamol,
purine, piperazine, piperidine, N-ethyl piperidine, polyamine resin
and the like. Particularly preferred organic bases are
isopropylamine, diethylamine, ethanolamine, trimethylamine,
dicyclohexylamine, choline and caffeine.
[0107] "Pharmaceutical composition" refers to a formulation formed
with a compound of the invention and a medium generally acceptable
in the art for the delivery of the biologically active compound to
a mammal e.g. humans. Such a medium includes all pharmaceutically
acceptable carriers, diluents or excipients.
[0108] "Therapeutically effective amount" refers to an amount of a
compound of the invention which, when administered to a mammal,
preferably a human, is sufficient to effect treatment (as defined
below) of the tumor and/or cancer in the mammal, preferably a
human. The amount of a compound of the invention which constitutes
a "therapeutically effective amount" will vary depending on the
compound, the condition and its severity, and the age of the mammal
to be treated, but can be determined routinely by one of ordinary
skill in the art having regard to his own knowledge and to this
disclosure.
[0109] "Treating" or "treatment", as used herein, covers the
treatment of relevant disease or condition in a mammal, preferably
a human, having the relevant disease or disorder, and includes:
[0110] (i) preventing the disease or condition from occurring in a
mammal, in particular, when such mammal is predisposed to the
condition but has not yet been diagnosed as having it;
[0111] (ii) inhibiting the disease or condition, i.e. arresting its
development; or
[0112] (iii) relieving the disease or condition, i.e. causing
regression of the disease or condition.
[0113] Throughout the treatment course, the administration in vivo
can be carried out by means of a single administration, a
continuous administration or an intermittent administration (such
as the administration is carried out by divided dose at appropriate
intervals). The method for determining the most effective
administration mode and dose would have been well-known for one of
ordinary skill in the art, and vary depending on the formulation to
be used in the treatment, the object of the treatment, the targeted
cell to be treated and the subject to be treated. A single or a
multiple administration can be carried out, and the level of dose
and the mode can be selected by an attending doctor.
SPECIFIC EMBODIMENTS
[0114] In one aspect, the present application relates to a compound
represented by formula (I) and a pharmaceutically acceptable salt
thereof,
##STR00082##
[0115] wherein:
[0116] R.sup.1 is selected from the group consisting of H,
optionally substituted alkyl, and optionally substituted
alkoxy;
[0117] R.sup.2 is selected from the group consisting of H,
optionally substituted aryl, optionally substituted heteroaryl,
optionally substituted (alkyleneoxy).sub.malkyl, optionally
substituted heterocyclyl, optionally substituted alkyl, and
optionally substituted sulfonyl;
[0118] R.sup.3 is selected from the group consisting of H,
optionally substituted aryl, optionally substituted heteroaryl,
optionally substituted (alkyleneoxy).sub.malkyl, optionally
substituted heterocyclyl, optionally substituted alkyl, and
optionally substituted sulfonyl;
[0119] or NR.sup.2R.sup.3 represents optionally substituted
heterocyclyl;
[0120] m is selected from the group consisting of 0, 1, 2, 3, 4, 5,
6, 7, 8, 9, 10, and 11; W is selected from the group consisting of
O and NH;
[0121] R.sup.4 is selected from the group consisting of H, F, and
optionally substituted alkyl;
[0122] R.sup.5 is selected from the group consisting of H, F,
optionally substituted alkyl and OR.sup.6, wherein R.sup.6 is
selected from the group consisting of H and tetrahydropyran-2-yl;
and
[0123] n is selected from the group consisting of 1, 2 and 3.
[0124] In another aspect, the present application relates to a
compound represented by formula (I) and a pharmaceutically
acceptable salt thereof,
##STR00083##
[0125] wherein:
[0126] R.sup.1 is selected from the group consisting of H,
C.sub.1-4alkyl, and C.sub.1-4alkoxy;
[0127] R.sup.2 is selected from the group consisting of H, aryl,
heteroaryl, (C.sub.1-4alkyleneoxy).sub.mC.sub.1-4alkyl,
heterocyclyl, C.sub.1-4alkyl, and sulfonyl;
[0128] R.sup.3 is selected from the group consisting of H, aryl,
heteroaryl, (C.sub.1-4alkyleneoxy).sub.mC.sub.1-4alkyl,
heterocyclyl, C.sub.1-4alkyl, and sulfonyl;
[0129] or NR.sup.2R.sup.3 represents heterocyclyl;
[0130] m is selected from the group consisting of 0, 1, 2, 3, 4, 5,
6, 7, 8, 9, 10, and 11;
[0131] W is selected from the group consisting of O and NH;
[0132] R.sup.4 is selected from the group consisting of H, F and
C.sub.1-4alkyl;
[0133] R.sup.5 is selected from the group consisting of H, F,
C.sub.1-4alkyl and OR.sup.6, wherein R.sup.6 is selected from the
group consisting of H and tetrahydropyran-2-yl; and
[0134] n is selected from the group consisting of 1, 2 and 3.
[0135] In some embodiments, R.sup.1 in the compound represented by
formula (I) is selected from the group consisting of H and
OCH.sub.3.
[0136] In some embodiments, W in the compound represented by
formula (I) is O.
[0137] In some embodiments, R.sup.4 in the compound represented by
formula (I) is CH.sub.3.
[0138] In some embodiments, R.sup.5 in the compound represented by
formula (I) is selected from the group consisting of OH and
(tetrahydropyran-2-yl)oxy.
[0139] In some embodiments, R.sup.2 in the compound represented by
formula (I) is selected from the group consisting of H, methyl,
ethyl, (morpholinylmethyl)phenyl, 4-((morpholin-1-yl)methyl)phenyl,
(dimethylaminomethyl)phenyl, 4-((dimethylamino)methyl)phenyl,
2-(2-(dimethylamino)ethoxy)ethyl, morpholin-1-yl, piperidin-1-yl,
tetrahydropyrrol-1-yl, 4-(2-hydroxyethyl)piperazin-1-yl,
(4-methylpiperazin)-1-yl, (4-ethylpiperazin)-1-yl,
2-(tetrahydropyrrol-1-yl)ethyl, 3-(tetrahydropyrrol-1-yl)propyl,
(2-(morpholin-1-yl)pyridin)-4-yl, (2-(morpholin-1-yl)pyridin)-5-yl,
pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, (pyridin-4-yl)methyl,
(pyridin-3-yl)methyl, (pyridin-2-yl)methyl, 2-(pyridin-4-yl)ethyl,
2-(pyridin-3-yl)ethyl, 2-(pyridin-2-yl)ethyl,
2-(pyridin-4-yl)propyl, 2-(pyridin-3-yl)propyl,
2-(pyridin-2-yl)propyl, 2-((4-sulfamido)phenyl)ethyl,
(3-(dimethylamino)propyl)piperazin-1-yl,
3-((4-sulfamido)phenyl)propyl, 3-((4-methyl)piperazin-1-yl)propyl,
3-((4-ethyl)piperazin-1-yl)propyl,
3-((4-propyl)piperazin-1-yl)propyl,
2-((4-methyl)piperazin-1-yl)ethyl,
2-((4-ethyl)piperazin-1-yl)ethyl,
2-((4-propyl)piperazin-1-yl)ethyl, 2-(dimethylamino)ethyl,
2-(diethylamino)ethyl.sub.2, 2-(dipropylamino)ethyl,
2-(piperidin-1-yl)ethyl, 2-(morpholin-1-yl)ethyl,
2-(tetrahydropyrrol-1-yl)ethyl, 3-(dimethylamino)propyl,
3-(diethylamino)propyl, 3-(dipropylamino)propyl,
3-(piperidin-1-yl)propyl, 3-(morpholin-1-yl)propyl,
3-(tetrahydropyrrol-1-yl)propyl, 4-(dimethylamino)butyl,
4-(diethylamino)butyl, 4-(dipropylamino)butyl,
4-(piperidin-1-yl)butyl, 4-(morpholin-1-yl)butyl,
4-(tetrahydropyrrol-1-yl)butyl,
2-(2-(2-(2-(dimethylamino)ethoxy)ethoxy)ethoxy)ethyl,
2-(2-(2-(2-(diethylamino)ethoxy)ethoxy)ethoxy)ethyl,
2-(2-(2-(2-(dipropylamino)ethoxy)ethoxy)ethoxy)ethyl,
2-(2-(2-(2-(piperidin-1-yl)ethoxy)ethoxy)ethoxy)ethyl,
2-(2-(2-(2-(morpholin-1-yl)ethoxy)ethoxy)ethoxy)ethyl,
2-(2-(2-(2-(tetrahydropyrrol-1-yl)ethoxy)ethoxy)ethoxy)ethyl,
2-(2-(2-(dimethylamino)ethoxy)ethoxy)ethyl,
2-(2-(2-(diethylamino)ethoxy)ethoxy)ethyl,
2-(2-(2-(dipropylamino)ethoxy)ethoxy)ethyl,
2-(2-(2-(piperidin-1-yl)ethoxy)ethoxy)ethyl,
2-(2-(2-(morpholin-1-yl)ethoxy)ethoxy)ethyl,
2-(2-(2-(tetrahydropyrrol-1-yl)ethoxy)ethoxy)ethyl, 6-purinyl,
mesyl, benzenesulfonyl, pyrazin-2-yl, pyrimidin-2-yl,
2-hydroxyethyl, 2-(2-hydroxyethoxy)ethyl,
2-(2-(2-(2-(2-(2-(2-(2-(2-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)ethoxy)eth-
oxy)ethoxy)ethoxy)ethoxy)ethoxy)ethoxy)ethoxy)ethyl,
2-(2-(2-(2-(2-(2-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)ethoxy)ethoxy)ethox-
y)ethoxy)ethoxy)ethyl,
2-(2-(2-(2-(2-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)ethoxy)ethoxy)ethoxy)e-
thoxy)ethyl,
2-(2-(2-(2-(2-(2-(2-(2-methoxy)ethoxy)ethoxy)ethoxy)ethoxy)ethoxy)ethoxy)-
ethoxy)ethoxy)ethyl, and
2-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)ethyl.
[0140] In some embodiments, R.sup.3 in the compound represented by
formula (I) is selected from the group consisting of H, methyl,
ethyl, (morpholinylmethyl)phenyl, 4-((morpholin-1-yl)methyl)phenyl,
(dimethylaminomethyl)phenyl, 4-((dimethylamino)methyl)phenyl,
2-(2-(dimethylamino)ethoxy)ethyl, morpholin-1-yl, piperidin-1-yl,
tetrahydropyrrol-1-yl, 4-(2-hydroxyethyl)piperazin-1-yl,
(4-methylpiperazin)-1-yl, (4-ethylpiperazin)-1-yl,
2-(tetrahydropyrrol-1-yl)ethyl, 3-(tetrahydropyrrol-1-yl)propyl,
(2-(morpholin-1-yl)pyridin)-4-yl, (2-(morpholin-1-yl)pyridin)-5-yl,
pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, (pyridin-4-yl)methyl,
(pyridin-3-yl)methyl, (pyridin-2-yl)methyl, 2-(pyridin-4-yl)ethyl,
2-(pyridin-3-yl)ethyl, 2-(pyridin-2-yl)ethyl,
2-(pyridin-4-yl)propyl, 2-(pyridin-3-yl)propyl,
2-(pyridin-2-yl)propyl, 2-((4-sulfamido)phenyl)ethyl,
(3-(dimethylamino)propyl)piperazin-1-yl,
3-((4-sulfamido)phenyl)propyl, 3-((4-methyl)piperazin-1-yl)propyl,
3-((4-ethyl)piperazin-1-yl)propyl,
3-((4-propyl)piperazin-1-yl)propyl,
2-((4-methyl)piperazin-1-yl)ethyl,
2-((4-ethyl)piperazin-1-yl)ethyl,
2-((4-propyl)piperazin-1-yl)ethyl, 2-(dimethylamino)ethyl,
2-(diethylamino)ethyl.sub.2, 2-(dipropylamino)ethyl,
2-(piperidin-1-yl)ethyl, 2-(morpholin-1-yl)ethyl,
2-(tetrahydropyrrol-1-yl)ethyl, 3-(dimethylamino)propyl,
3-(diethylamino)propyl, 3-(dipropylamino)propyl,
3-(piperidin-1-yl)propyl, 3-(morpholin-1-yl)propyl,
3-(tetrahydropyrrol-1-yl)propyl, 4-(dimethylamino)butyl,
4-(diethylamino)butyl, 4-(dipropylamino)butyl,
4-(piperidin-1-yl)butyl, 4-(morpholin-1-yl)butyl,
4-(tetrahydropyrrol-1-yl)butyl,
2-(2-(2-(2-(dimethylamino)ethoxy)ethoxy)ethoxy)ethyl,
2-(2-(2-(2-(diethylamino)ethoxy)ethoxy)ethoxy)ethyl,
2-(2-(2-(2-(dipropylamino)ethoxy)ethoxy)ethoxy)ethyl,
2-(2-(2-(2-(piperidin-1-yl)ethoxy)ethoxy)ethoxy)ethyl,
2-(2-(2-(2-(morpholin-1-yl)ethoxy)ethoxy)ethoxy)ethyl,
2-(2-(2-(2-(tetrahydropyrrol-1-yl)ethoxy)ethoxy)ethoxy)ethyl,
2-(2-(2-(dimethylamino)ethoxy)ethoxy)ethyl,
2-(2-(2-(diethylamino)ethoxy)ethoxy)ethyl,
2-(2-(2-(dipropylamino)ethoxy)ethoxy)ethyl,
2-(2-(2-(piperidin-1-yl)ethoxy)ethoxy)ethyl,
2-(2-(2-(morpholin-1-yl)ethoxy)ethoxy)ethyl,
2-(2-(2-(tetrahydropyrrol-1-yl)ethoxy)ethoxy)ethyl, 6-purinyl,
mesyl, benzenesulfonyl, pyrazin-2-yl, pyrimidin-2-yl,
2-hydroxyethyl, 2-(2-hydroxyethoxy)ethyl,
2-(2-(2-(2-(2-(2-(2-(2-(2-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)ethoxy)eth-
oxy)ethoxy)ethoxy)ethoxy)ethoxy)ethoxy)ethoxy)ethyl,
2-(2-(2-(2-(2-(2-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)ethoxy)ethoxy)ethox-
y)ethoxy)ethoxy)ethyl,
2-(2-(2-(2-(2-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)ethoxy)ethoxy)ethoxy)e-
thoxy)ethyl,
2-(2-(2-(2-(2-(2-(2-(2-methoxy)ethoxy)ethoxy)ethoxy)ethoxy)ethoxy)ethoxy)-
ethoxy)ethoxy)ethyl, and
2-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)ethyl.
[0141] In some embodiments, NR.sup.2R.sup.3 in the compound
represented by formula (I) is selected from the group consisting of
piperidin-1-yl, morpholin-1-yl, tetrahydropyrrol-1-yl,
(4-(2-hydroxyethyl))piperazin-1-yl, (4-methyl)piperazin-1-yl,
(4-ethyl)piperazin-1-yl, (4-propyl)piperazin-1-yl,
4-(3-hydroxypropyl)piperazin-1-yl, 3-(morpholin-1-yl)propyl,
adenin-1-yl, (4-(3-(dimethylamino)propyl)piperazin)-1-yl,
(4-(2-(dimethylamino)ethyl)piperazin)-1-yl,
(4-(3-(diethylamino)propyl)piperazin)-1-yl,
(4-(2-(diethylamino)ethyl)piperazin)-1-yl,
(4-(2-(piperidin-1-yl)ethyl)piperazin)-1-yl,
(4-(3-(piperidin-1-yl)propyl)piperazin)-1-yl,
(4-(2-(morpholin-1-yl)ethyl)piperazin)-1-yl,
(4-(3-(morpholin-1-yl)propyl)piperazin)-1-yl,
(4-(2-(tetrahydropyrrol-1-yl)ethyl)piperazin)-1-yl, and
(4-(3-(tetrahydropyrrol-1-yl)propyl)piperazin)-1-yl.
[0142] In still another aspect, the present application relates to
compounds selected from the group consisting of:
TABLE-US-00002 ##STR00084## No. n NR.sup.2R.sup.3 1 3 ##STR00085##
2 2 ##STR00086## 3 3 ##STR00087## 4 2 ##STR00088## 5 3 ##STR00089##
6 2 ##STR00090## 7 3 ##STR00091## 8 2 ##STR00092## 9 3 ##STR00093##
10 2 ##STR00094## 11 3 ##STR00095## 12 2 ##STR00096## 13 3
##STR00097## 14 2 ##STR00098## 15 3 ##STR00099## 16 2 ##STR00100##
17 3 ##STR00101## 18 2 ##STR00102## 19 3 ##STR00103## 20 2
##STR00104## 21 3 ##STR00105## 22 2 ##STR00106## 23 3 ##STR00107##
24 2 ##STR00108## 25 3 ##STR00109## 26 2 ##STR00110## 27 3
##STR00111## 28 2 ##STR00112## 29 3 ##STR00113## 30 2 ##STR00114##
31 3 ##STR00115## 32 2 ##STR00116## 33 3 ##STR00117## 34 2
##STR00118## 35 3 ##STR00119## 36 2 ##STR00120## 37 3 ##STR00121##
38 2 ##STR00122## 39 3 ##STR00123## 40 2 ##STR00124## 41 3
##STR00125## 42 2 ##STR00126## 43 3 ##STR00127## 44 2 ##STR00128##
45 3 ##STR00129## 46 2 ##STR00130## 47 3 ##STR00131## 48 2
##STR00132## 49 3 ##STR00133## 50 2 ##STR00134## 51 3 ##STR00135##
52 2 ##STR00136## 53 3 ##STR00137## 54 2 ##STR00138## 55 3
##STR00139## 56 2 ##STR00140## 57 3 ##STR00141## 58 2 ##STR00142##
59 3 ##STR00143## 60 2 ##STR00144## 61 3 ##STR00145## 62 2
##STR00146## 63 3 ##STR00147## 64 2 ##STR00148## 65 3 ##STR00149##
66 2 NH.sub.2 67 3 NH.sub.2 68 2 NHCH.sub.3 69 3 NHCH.sub.3 70 2
N(CH.sub.3).sub.2 71 3 N(CH.sub.3).sub.2 72 2 ##STR00150## 73 3
##STR00151## 74 2 ##STR00152## 75 2 ##STR00153## 88 2 ##STR00154##
90 2 ##STR00155## 91 2 ##STR00156## 92 2 ##STR00157## 93 2
##STR00158## 94 2 ##STR00159## 95 2 ##STR00160##
[0143] 77)
10-((3'-(pyrrol-1-yl)-2',3',6'-trideoxy-alphal-L-lyxo-hexylpyranyl)oxy)-7-
,8,9,10-tetrahydro-6,8,11-trihydroxy-13-oxo-14-(4-((4-(2-hydroxy)ethyl)pip-
erazin-1-yl)-4-oxo-butyrato)-1-methoxy-5,12-naphthalenedione
acetate; [0144] 78)
10-((3'-(pyrrol-1-yl)-2',3',6'-trideoxy-alphal-L-lyxo-hexylpyranyl)oxy)-7-
,8,9,10-tetrahydro-6,8,11-trihydroxy-13-oxo-14-(4-((3-(morpholin-1-yl)prop-
yl)amino)-4-oxo-butyrato)-1-m ethoxy-5,12-naphthalenedione acetate;
[0145] 79)
10-((3'-(pyrrol-1-yl)-2',3',6'-trideoxy-alphal-L-lyxo-hexylpyranyl)ox-
y)-7,8,9,10-tetrahydro-6,8,11-trihydroxy-13-oxo-14-(4-((4-methyl)piperazin-
-1-yl)amino)-4-oxo-butyrato)-1-methoxy-5,12-naphthalenedione
acetate; [0146] 80)
10-((3'-(pyrrol-1-yl)-2',3',6'-trideoxy-alphal-L-lyxo-hexylpyranyl)oxy)-7-
,8,9,10-tetrahydro-6,8,11-trihydroxy-13-oxo-14-(4-(4-ethylpiperazin-1-yl)--
4-oxo-butyrato)-1-methoxy-5,12-naphthalenedione acetate; [0147] 81)
10-((3'-(pyrrol-1-yl)-2',3',6'-trideoxy-alphal-L-lyxo-hexylpyranyl)oxy)-7-
,8,9,10-tetrahydro-6,8,11-trihydroxy-13-oxo-14-(4-((3-(4-methylpiperazin-1-
-yl)propyl)amino)-4-oxo-butyrato)-1-methoxy-5,12-naphthalenedione
acetate; [0148] 82)
10-((3'-(pyrrol-1-yl)-2',3',6'-trideoxy-alphal-L-lyxo-hexylpyranyl)oxy)-7-
,8,9,10-tetrahydro-6,8,11-trihydroxy-13-oxo-14-(4-((pyridin-4-yl)methyl)am-
ino)-4-oxo-butyrato)-1-methoxy-5,12-naphthalenedione acetate;
[0149] 83)
10-((3'-(pyrrol-1-yl)-2',3',6'-trideoxy-alphal-L-lyxo-hexylpyranyl)oxy)-7-
,8,9,10-tetrahydro-6,8,11-trihydroxy-13-oxo-14-(4-((pyridin-3-yl)methyl)am-
ino)-4-oxo-butyrato)-1-methoxy-5,12-naphthalenedione acetate;
[0150] 84)
10-((3'-(pyrrol-1-yl)-2',3',6'-trideoxy-alphal-L-lyxo-hexylpyranyl)oxy)-7-
,8,9,10-tetrahydro-6,8,11-trihydroxy-13-oxo-14-(4-(2-(pyridin-2-yl)ethyl)a-
mino)-4-oxo-butyrato)-1-methoxy-5,12-naphthalenedione acetate;
[0151] 85)
10-((3'-(pyrrol-1-yl)-2',3',6'-trideoxy-alphal-L-lyxo-hexylpyranyl)oxy)-7-
,8,9,10-tetrahydro-6,8,11-trihydroxy-13-oxo-14-(4-(2-(pyridin-3-yl)ethyl)a-
mino)-4-oxo-butyrato)-1-methoxy-5,12-naphthalenedione acetate;
[0152] 86)
10-((3'-(pyrrol-1-yl)-2',3',6'-trideoxy-alphal-L-lyxo-hexylpyranyl)oxy)-7-
,8,9,10-tetrahydro-6,8,11-trihydroxy-13-oxo-14-(4-(2-(pyridin-4-yl)ethyl)a-
mino)-4-oxo-butyrato)-1-methoxy-5,12-naphthalenedione acetate;
[0153] 87)
10-((3'-(pyrrol-1-yl)-2',3',6'-trideoxy-alphal-L-lyxo-hexylpyranyl)oxy)-7-
,8,9,10-tetrahydro-6,8,11-trihydroxy-13-oxo-14-(4-(4-(3-(dimethylamino)pro-
pyl)piperazin-1-yl)-4-oxo-butyrato)-1-methoxy-5,12-naphthalenedione
acetate; [0154] 89)
10-((3'-(pyrrol-1-yl)-2',3',6'-trideoxy-alphal-L-lyxo-hexylpyranyl)oxy)-7-
,8,9,10-tetrahydro-6,8,11-trihydroxy-13-oxo-14-(4-((2-(morpholin-1-yl)ethy-
l)amino)-4-oxo-butyrato)-1-methoxy-5,12-naphthalenedione acetate;
[0155] 96)
10-((3'-(pyrrol-1-yl)-2',3',6'-trideoxy-alphal-L-lyxo-hexylpyranyl)ox-
y)-7,8,9,10-tetrahydro-6,8,11-trihydroxy-13-oxo-14-(4-((2-(morpholin-1-yl)-
ethyl)amino)-4-oxo-butyrato)-1-methoxy-5,12-naphthalenedione
phosphate; and [0156] 99)
10-((3'-(pyrrol-1-yl)-2',3',6'-trideoxy-alphal-L-lyxo-hexylpyranyl)oxy)-7-
,8,9,10-tetrahydro-6,8,11-trihydroxy-13-oxo-14-(4-((2-(morpholin-1-yl)ethy-
l)amino)-4-oxo-butyrato)-1-methoxy-5,12-naphthalenedione
hydrochloride.
[0157] In yet still another aspect, the present application relates
to a process for preparing a compound represented by formula (I),
comprising:
##STR00161##
[0158] reacting a compound represented by formula (II) with a
compound represented by formula (III) in the presence of a
condensation agent to obtain the compound represented by formula
(I),
[0159] wherein:
[0160] in the compound represented by formula (I),
[0161] R.sup.1 is selected from the group consisting of H,
C.sub.1-4alkyl, and C.sub.1-4alkoxy; R.sup.2 is selected from the
group consisting of H, aryl, heteroaryl,
(C.sub.1-4alkyleneoxy).sub.mC.sub.1-4alkyl, heterocyclyl,
C.sub.1-4alkyl, and sulfonyl; R.sup.3 is selected from the group
consisting of H, aryl, heteroaryl,
(C.sub.1-4alkyleneoxy).sub.mC.sub.1-4alkyl, heterocyclyl,
C.sub.1-4alkyl, and sulfonyl; or NR.sup.2R.sup.3 represents
heterocyclyl; m is selected from the group consisting of 0, 1, 2,
3, 4, 5, 6, 7, 8, 9, 10, and 11; W is selected from the group
consisting of O and NH; R.sup.4 is selected from the group
consisting of H, F and C.sub.1-4alkyl; R.sup.5 is selected from the
group consisting of H, F, C.sub.1-4alkyl and OR.sup.6, in which
R.sup.6 is selected from the group consisting of H and
tetrahydropyran-2-yl; n is selected from the group consisting of 1,
2 and 3;
[0162] groups represented by R.sup.1, W, R.sup.4, R.sup.5 in the
compound represented by formula (II) are the same as groups
represented by R.sup.1, W, R.sup.4, R.sup.5 in the compound
represented by formula (I);
[0163] n in the compound represented by formula (III) has the same
meanings as n in the compound represented by formula (I); groups
represented by R.sup.7 and R.sup.8 in the compound represented by
formula (III) are the same as groups represented by R.sup.2 and
R.sup.3 in the compound represented by formula (I), provided that
groups represented by R.sup.7 and R.sup.8 do not comprise NH or
NH.sub.2; when groups represented by R.sup.7 and R.sup.8 comprise
NH or NH.sub.2, the compound represented by formula (III) has an
amino-protecting group at N-terminus, and is subject to a
deprotection reaction to obtain the compound represented by formula
(I).
[0164] In some embodiments, the process for preparing a compound
represented by formula (I) further comprises adding an
activator.
[0165] Exemplary examples of activators that can be used in the
process for preparing a compound represented by formula (I)
according to the present application include, but are not limited
to, N-hydroxysuccinimide (HOSu), 1-hydroxy-7-azobenzotriazole
(HOAt), 1-hydroxybenzotriazole (HOBt), N-hydroxyphthalimide (NHPI),
N-hydroxy-1,8-naphthalimide (NHNI), pentafluorophenol (PFPOH),
2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium
hexafluorophosphate (HATU),
benzotriazole-N,N,N',N'-tetramethyluronium hexafluorophosphate
(HBTU), 6-chlorobenzotriazole-1,1,3,3-tetramethyluronium
hexafluorophosphate (HCTU),
O-(7-azabenzotriazole-1-yl)-di(tetrahydropyrrolyl)carbenium
hexafluophosphate (HAPyU),
O-(benzotriazole-1-yl)-di(tetrahydropyrrolyl)carbenium
hexafluophosphate (HBPyU),
O-benzotriazole-N,N,N',N'-tetramethyluronium tetrafluoroborate
(TBTU), 2-succinimido-1,1,3,3-tetramethyluronium tetrafluoroborate
(TSTU), 2-(5-norbomen-2,3-dicarboximido)-1,1,3,3-tetramethyluronium
tetrafluoroborate quaternary ammonium salt (TNTU),
benzotriazole-1-yloxytri(dimethylamino)phosphonium
hexafluorophosphate (BOP),
benzotriazol-1-yl-oxytripyrrolidino-phosphonium hexafluorophosphate
(PyBOP),
(3H-1,2,3-triazolo[4,5-b]pyridin-3-oxy)tri-1-pyrrolidinylphosphonium
hexafluorophosphate (PyAOP), diphenylphosphinyl chloride (DPP-Cl),
diphenyl phosphoryl azide (DPPA), cyanodiethylphosphate (DECP),
bis(2-oxo-3-oxazolidinyl)phosphinyl chloride (BOP-Cl) and a mixture
thereof.
[0166] Exemplary examples of condensation agents that can be used
in the process for preparing a compound represented by formula (I)
according to the present application include, but are not limited
to, dicyclohexylcarbodiimide (DCC), diisopropylcarbodiimide (DIC),
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDCI),
2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium
hexafluorophosphate (HATU),
benzotriazol-N,N,N',N'-tetramethyluronium hexafluorophosphate
(HBTU), 6-chlorobenzotriazol-1,1,3,3-tetramethyluronium
hexafluorophosphate (HCTU),
O-(7-azabenzotriazol-1-yl)-di(tetrahydropyrrolyl)carbenium
hexafluophosphate (HAPyU),
O-(benzotriazol-1-yl)-di(tetrahydropyrrolyl)carbenium
hexafluophosphate (HBPyU),
O-benzotriazol-N,N,N',N'-tetramethyluronium tetrafluoroborate
(TBTU), 2-succinimido-1,1,3,3-tetramethyluronium tetrafluoroborate
(TSTU), 2-(5-norbomen-2,3-dicarboximido)-1,1,3,3-tetramethyluronium
tetrafluoroborate quaternary ammonium salt (TNTU),
benzotriazol-1-yloxytri(dimethylamino)phosphonium
hexafluorophosphate (BOP),
benzotriazol-1-yl-oxytripyrrolidino-phosphonium hexafluorophosphate
(PyBOP),
(3H-1,2,3-triazolo[4,5-b]pyridin-3-oxy)tri-1-pyrrolidinylphosphonium
hexafluorophosphate (PyAOP), diphenylphosphinyl chloride (DPP-Cl),
diphenyl phosphoryl azide (DPPA), cyanodiethylphosphate (DECP),
bis(2-oxo-3-oxazolidinyl)phosphinyl chloride (BOP-Cl) and a mixture
thereof.
[0167] In some embodiments, the process for preparing a compound
represented by formula (I) further comprises adding a catalyst.
[0168] Exemplary examples of catalysts that can be used in the
process for preparing a compound represented by formula (I)
according to the present application include, but are not limited
to, 4-dimethylaminopyridine, 4-pyrrolidinylpyridine and a mixture
thereof.
[0169] Exemplary examples of nitrogen-protecting groups that can be
used in the process for preparing a compound represented by formula
(I) according to the present application include, but are not
limited to, Fmoc (fluorenylmethoxycarbony), Boc
(t-butyloxycarboryl), CBZ (carbobenzoxy), Tr (trityl) or Alloc
(allyloxycarbonyl), Teoc (trimethylsilylethoxycarbonyl),
methoxycarbonyl, ethoxycarbonyl, Pht (phthaloyl), Tos (tosyl), Ns
(o/p-nitrobenzenesulfonyl), Tfa (trifluoroacetyl), pivaloyl,
benzoyl, Trt (trityl), Dmb (2,4-dimethoxybenzyl), PMB
(p-methoxybenzyl), and Bn (benzyl).
[0170] Exemplary examples of deprotection reactants that can be
used in the process for preparing a compound represented by formula
(I) according to the present application include, but are not
limited to, hydrogen gas, NH.sub.3, aminoethanol, dimethylamine,
diethylamine, piperidine, piperazine, DBU, hydrochloric acid,
phosphoric acid, acetic acid, formic acid, trifluoroacetic acid,
methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid
and a mixture thereof.
[0171] In some embodiments, the compound represented by formula
(III) in the process for preparing a compound represented by
formula (I) is obtained by a reaction of a compound represented by
formula (IV) with HNR.sup.7R.sup.8,
##STR00162##
[0172] wherein:
[0173] n in the compound represented by formula (IV) has the same
meanings as n in the compound represented by formula (I);
[0174] groups represented by R.sup.2 and R.sup.3 in
HNR.sup.7R.sup.8 are the same as groups represented by R.sup.2 and
R.sup.3 in the compound represented by formula (I).
[0175] In some embodiments, the process for preparing a compound
represented by formula (III) further comprises adding an alkaline
compound.
[0176] Exemplary examples of alkaline compounds that can be used in
the process for preparing a compound represented by formula (III)
according to the present application include, but are not limited
to, triethylamine, pyridine, diisopropylethylamine, trimethylamine,
N-methylpyrrolidine, N-methylpiperidine, N-methylmorpholine,
N-ethylpyrrolidine, N-ethylpiperidine, N-ethylmorpholine and a
mixture thereof.
[0177] In yet another aspect, the present application relates to a
pharmaceutical composition comprising a compound represented by
formula (I) or a pharmaceutically acceptable salt thereof, and a
pharmaceutically acceptable carrier,
##STR00163##
[0178] wherein:
[0179] R.sup.1 is selected from the group consisting of H,
C.sub.1-4alkyl, and C.sub.1-4alkoxy;
[0180] R.sup.2 is selected from the group consisting of H, aryl,
heteroaryl, (C.sub.1-4alkyleneoxy).sub.mC.sub.1-4alkyl,
heterocyclyl, C.sub.1-4alkyl, and sulfonyl;
[0181] R.sup.3 is selected from the group consisting of H, aryl,
heteroaryl, (C.sub.1-4alkyleneoxy).sub.mC.sub.1-4alkyl,
heterocyclyl, C.sub.1-4alkyl, and sulfonyl;
[0182] or NR.sup.2R.sup.3 represents heterocyclyl;
[0183] m is selected from the group consisting of 0, 1, 2, 3, 4, 5,
6, 7, 8, 9, 10, and 11;
[0184] W is selected from the group consisting of O and NH;
[0185] R.sup.4 is selected from the group consisting of H, F and
C.sub.1-4alkyl;
[0186] R.sup.5 is selected from the group consisting of H, F,
C.sub.1-4alkyl and OR.sup.6, wherein R.sup.6 is selected from the
group consisting of H and tetrahydropyran-2-yl; and
[0187] n is selected from the group consisting of 1, 2 and 3.
[0188] In another aspect, the present application relates to a
formulation comprising a compound represented by formula (I) or a
pharmaceutically acceptable salt thereof, and a pharmaceutically
acceptable carrier,
##STR00164##
[0189] wherein:
[0190] R.sup.1 is selected from the group consisting of H,
C.sub.1-4alkyl, and C.sub.1-4alkoxy;
[0191] R.sup.2 is selected from the group consisting of H, aryl,
heteroaryl, (C.sub.1-4alkyleneoxy).sub.mC.sub.1-4alkyl,
heterocyclyl, C.sub.1-4alkyl, and sulfonyl;
[0192] R.sup.3 is selected from the group consisting of H, aryl,
heteroaryl, (C.sub.1-4alkyleneoxy).sub.mC.sub.1-4alkyl,
heterocyclyl, C.sub.1-4alkyl, and sulfonyl;
[0193] or NR.sup.2R.sup.3 represents heterocyclyl;
[0194] m is selected from the group consisting of 0, 1, 2, 3, 4, 5,
6, 7, 8, 9, 10, and 11;
[0195] W is selected from the group consisting of O and NH;
[0196] R.sup.4 is selected from the group consisting of H, F and
C.sub.1-4alkyl;
[0197] R.sup.5 is selected from the group consisting of H, F,
C.sub.1-4alkyl and OR.sup.6, wherein R.sup.6 is selected from the
group consisting of H and tetrahydropyran-2-yl; and
[0198] n is selected from the group consisting of 1, 2 and 3.
[0199] In some embodiments, the formulation comprising a compound
represented by formula (I) or a pharmaceutically acceptable salt
thereof and a pharmaceutically acceptable carrier is a formulation
for injection.
[0200] Exemplary examples that can be used in the formulation
according to the present application include, but are not limited
to, a conventional powder injection, a freeze-dried powder
injection, a hydro-injection, an emulsion, a solution and a
suspension.
[0201] In still another aspect, the present application relates to
a method for treating and/or preventing tumor and/or cancer,
comprising administering to a subject in need thereof a
therapeutically effective amount of a compound represented by
formula (I) or a pharmaceutically acceptable salt thereof, or
administering a therapeutically effective amount of a
pharmaceutical composition comprising a compound represented by
formula (I) or a pharmaceutically acceptable salt and a
pharmaceutically acceptable carrier, or administering a
therapeutically effective amount of a formulation comprising a
compound represented by formula (I) or a pharmaceutically
acceptable salt and a pharmaceutically acceptable carrier,
##STR00165##
[0202] wherein:
[0203] R.sup.1 is selected from the group consisting of H,
C.sub.1-4alkyl, and C.sub.1-4alkoxy;
[0204] R.sup.2 is selected from the group consisting of H, aryl,
heteroaryl, (C.sub.1-4alkyleneoxy).sub.mC.sub.1-4alkyl,
heterocyclyl, C.sub.1-4alkyl, and sulfonyl;
[0205] R.sup.3 is selected from the group consisting of H, aryl,
heteroaryl, (C.sub.1-4alkyleneoxy).sub.mC.sub.1-4alkyl,
heterocyclyl, C.sub.1-4alkyl, and sulfonyl;
[0206] or NR.sup.2R.sup.3 represents heterocyclyl;
[0207] m is selected from the group consisting of 0, 1, 2, 3, 4, 5,
6, 7, 8, 9, 10, and 11;
[0208] W is selected from the group consisting of O and NH;
[0209] R.sup.4 is selected from the group consisting of H, F and
C.sub.1-4alkyl;
[0210] R.sup.5 is selected from the group consisting of H, F,
C.sub.1-4alkyl and OR.sup.6, wherein R.sup.6 is selected from the
group consisting of H and tetrahydropyran-2-yl; and
[0211] n is selected from the group consisting of 1, 2 and 3.
[0212] Exemplary examples of tumors and/or cancers that can be
treated and/or prevented by the method according to the present
application include, but are not limited to, liver cancer, gastric
cancer, breast cancer, lung cancer, intestine cancer, ovarian
cancer, pancreatic cancer, head and neck cancer, cervical cancer,
renal cancer, melanoma, prostatic cancer, brain glioma, various
leukemia, lymphoma, and multiple bone marrow cancer.
[0213] The compound and a salt thereof according to the present
application possess good anticancer and/or antitumor activity, and
good water solubility and stability, as well as good tolerance in
animal bodies. Therefore, they are prone to being developed as
clinical drugs.
EXAMPLES
[0214] Although any one skilled in the art is capable of preparing
the compounds of the present application according to the general
techniques disclosed herein above, more specific details on
synthetic techniques for the compound of the present application
are provided elsewhere in this specification for conveniences. In
addition, all reagents and reaction conditions employed in
synthesis are known to those skilled in the art and are available
from ordinary commercial sources.
Abbreviations
[0215] Su: succinimide; Bt: benzotrazol-1-yl; At:
7-azobenzotrazol-1-yl; Fmoc: fluorenylmethoxycarbonyl; Boc:
t-butoxycarbonyl; CBZ: carbobenzoxy; Tr: trimethylphenyl; Alloc:
allyloxycarbonyl; DBU: 1,8-diazacyclo[5,4,0]hendecene-7; DIEA:
diisopropylethylamine; DMAP: 4-dimethylaminopyridine; EDC-HCl:
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride; HOAt:
N-hydroxy-7-azobenzotriazole; DCC: dicyclohexylcarbodiimide; DIC:
N,N-diisopropylcarbodiimide. NCI-H446: human small cell lung cancer
cell line; BxPC-3: human pancreatic cancer cell line; SK-OV-3:
human ovarian cancer cell line; MDA-MB-453: human breast cancer
cell line; 22Rv1: human prostate cancer cell line; A375: human
cutaneous melanoma cell line; A431: human epidermal carcinoma cell
line; MCF-7: human breast cancer cell line; NCI-446: human small
cell lung cancer cell line; NCI-H460: human large cell lung cancer
cell line; B16: mouse melanoma cell line; 786-O: human kidney clear
cell adenocarcinoma cell line; DU-145: prostate cancer cell line;
Hep3B: liver cancer cell line; SK-Br-3: human breast cancer cell
line; MTT: nitroblue tetrazolium; McCoy's 5A: McCoy's 5A Medium;
FBS: fetal calf serum; PBS: phosphate buffer, pH 7.4; EDTA:
ethylenediamine tetraacetic acid; DMSO: dimethyl sulfoxide;
RPMI-1640: RPMI-1640 Medium; SRB: sulforhodamine; TCA:
trichloroacetic acid; ddH.sub.2O: double distilled water; Tris:
trihydroxymethylaminomethane; L15: Leibovitz's L-15 Medium;
compound A: 3'-pyrrolyldoxorubicin-14-oxo-succinic acid
monoester.
[0216] The numberings of the substituents in the present
application are indicated as follows.
##STR00166##
Preparation 1
3'-PYRROLYLDOXORUBICIN
10-((3'-(PYRROL-1-YL)-2',3',6'-TRIDEOXY-ALPHAL-L-LYXO-HEXYLPYRANYL)
OXY)-7,
8,9,10-TETRAHYDRO-6,8,11-TRIHYDROXY-8-HYDROXYACETYL-1-METHOXY-5,1-
2-NAPHTHALENEDIONE
[0217] To a three-neck flask (1 L) were added doxorubicin
hydrochloride (3.076 g), distilled water (300 mL) and
1,2-dichloroethane (300 mL), 2,5-dimethoxytetrahydrofuran (30 mL)
and glacial acetic acid (6 mL). The mixture was heated and refluxed
for 45 min under the protection of argon gas until the reaction was
completed. The reaction was cooled to the room temperature. The
reaction solution was poured into ice water (200 mL), and then
stood to separate. The organic phase was washed with saturated
saline (200 mL) once, dried over anhydrous magnesium sulfate,
filtered and rotary-evaporated to dryness. To the aqueous phase was
added 5% sodium bicarbonate aqueous solution (100 mL) while being
stirred in an ice bath, and then extracted with chloroform (50
mL.times.3). The chloroform layers were combined and washed with
saturated saline (100 mL) once, filtered and rotary-evaporated to
remove the solvent. The resultant crude was combined with the crude
obtained above. The resultant mixture was purified with column
chromatography, eluting with chloroform:methanol=35:1, to give the
product (2.91 g). MS: 592 (M-1)
Preparation 2
4-(4-NITROBENZYL)MORPHOLINE
[0218] To a reaction flask was added p-nitrobenzyl bromide (72.084
g). Dichloromethane (470 mL, dried via molecular sieve) was added
to dissolve p-nitrobenzyl bromide. Anhydrous potassium carbonate
(91.909 g) was added. The mixture was cooled in an ice bath under
the protection of argon gas. After 20 min, to the reaction flask
was dropwise added morpholine (over about 30 min). After the
addition was completed, the ice bath was removed. The resultant
mixture was stirred overnight at the room temperature. After the
reaction was completed, water (150 mL) was added. The pH of the
mixture was adjusted with 5% citric acid aqueous solution to 4-5
under stirring. After standing to separate, the organic phase was
washed with water (260 mL.times.1), dried over anhydrous MgSO.sub.4
for half an hour, and then filtered. The filtrate was concentrated,
and dried under reduced pressure by oil pump, to give the target
compound (71.9 g, yield 97.16%).
Preparation 3
4-(MORPHOLINYLMETHYL)ANILINE
[0219] To a reaction flask were added 4-(4-nitrobenzyl)morpholine
(37.4 g) and absolute ethanol (520 mL). The mixture was
mechanically stirred, and then acetic acid (34 mL) was added. The
resultant mixture was warmed in an oil bath. The solution became
clear at 40.degree. C. To 1 N hydrochloric acid (HCl) (84 mL) was
added iron powders (48.002 g), and stirred for 10 min, filtered by
suction. The filter cake was washed with absolute ethanol, and then
added into the reaction flask. The mixture was warmed in an oil
bath to reflux, and maintained under reflux until the reaction was
completed (about 3 hr). After the reaction was completed, the
reaction solution was filtered by suction. The filtrate was
concentrated, and then dissolved in ethyl acetate (400 mL) and
water (400 mL). The mixture was mixed and then stood to separate.
The organic layer was discarded. The aqueous phase was washed with
dichloromethane (100 mL.times.3). The pH was adjusted with NaOH
solids to 9. Lots of brown solids precipitated from the solution,
The solids were filtered by suction. The filter cake was washed
with distilled water (20 mL.times.2) and then discarded. The pH of
the filtrate was adjusted with sodium hydroxide (NaOH) to 13. The
resultant solution was extracted with dichloromethane (100
mL.times.2). The organic phase was directly concentrated to give
the target compound.
Preparation 4
5-(4-(MORPHOLINYLMETHYL)PHENYLAMINO)-5-OXOPENTANOIC ACID
[0220] To a reaction flask was added 4-(morpholinylmethyl)aniline
(960 mg). Dichloromethane (7.6 mL, dried via molecular sieve) was
added to dissolve 4-(morpholinylmethyl)aniline. The solution was
stirred under the protection of argon gas. To the reaction flask
were added glutaric anhydride (741 mg), DIEA (1.1 mL) and DMAP (62
mg). The mixture was stirred overnight at the room temperature.
After the reaction was completed, dichloromethane (50 mL) and
distilled water (30 mL) were added. The pH of the resultant mixture
was adjusted with NaOH solids to 13. The resultant solution was
mixed homogeneously again, and stood to separate. The pH of the
aqueous phase was adjusted with HCl to 3. The solution was frozen
to dry. The resultant solids were washed with absolute ethanol (50
mL) and filtered. The resultant filtrate was concentrated and
redissolved in dichloromethane. The resultant solution was
concentrated again to remove the residual absolute ethanol. The
resultant product was directly dried under reduced pressure to give
the target compound.
Preparation 5
N,N-DIMETHYL(4-NITROPHENYL)METHYLAMINE
[0221] To a reaction flask were added nitrobenzyl bromide (6.291 g)
and dichloromethane (50 mL). Anhydrous potassium carbonate (12.423
g) and dimethylamine hydrochloride (4.891 g) were successively
added to the reaction flask. The mixture was stirred overnight at
the room temperature. After the reaction was completed, the
reaction solution was filtered by suction. The filter cake was
washed with dichloromethane (10 mL). The filtrate was washed three
times with distilled water (50 mL.times.1, 30 mL.times.2). The
organic phase was directly concentrated and dried under reduced
pressure by oil pump to give an oil (4.553 g).
Preparation 6
4-((DIMETHYLAMINO)METHYL)ANILINE
[0222] To a reaction flask was added
N,N-dimethyl(4-nitrophenyl)methyamine (4.553 g). Anhydrous ethanol
was added to dissolve N,N-dimethyl(4-nitrophenyl)methyamine. After
adding acetic acid (5.2 mL), the mixture was mechanically stirred.
To 1 N HCl (40 mL) was added iron powders (11.339 g). The iron
powders were immersed for 10 min and filtered by suction. The
filter cake was washed with absolute ethanol and then added into
the reaction flask. The mixture was warmed in an oil bath to
reflux, and maintained under reflux until the reaction was
completed (about 35 min). After the reaction was completed, the
reaction solution was filtered by suction. The filter cake was
washed with absolute ethanol. The filtrate was concentrated and
then added into distilled water (100 mL). The pH of the mixture was
adjusted with NaOH to 14. Lots of solids were precipitated. The
solids were filtered by suction. The filtrate was extracted with
dichloromethane (100 mL.times.1), and the aqueous phase was
discarded. To the organic phase was added distilled water (60 mL).
The pH was adjusted with 2 N HCl to 2. The resultant product was
mixed and stood to separate. The organic phase was discarded. The
pH of the aqueous phase was adjusted with NaOH to 7. The mixture
was extracted with dichloromethane (40 mL.times.2) and the aqueous
phase was discarded. The organic phase was washed with distilled
water (40 mL.times.1) once and the aqueous phase was discarded. The
organic phase was directly concentrated to give the target
compound.
Preparation 7
T-BUTYL 2-(2-HYDROXYETHOXY)ETHYLCARBAMATE
[0223] To a single-neck flask was added 2-(2-aminoethoxy)ethanol
(10.500 g). Tetrahydrofuran (25 mL) was added to dissolve
2-(2-aminoethoxy)ethanol. Anhydrous sodium carbonate (5.300 g) was
dissolved in distilled water (30 mL). The solution was added into
the single-neck flask and cooled in an ice bath. Di-t-butyl
dicarbonate (28.340 g) was dissolved in tetrahydrofuran (70 mL).
The resultant solution was slowly dropwise added in the reaction
system (for about 1 hr). After the addition, the mixture was
stirred for 1.5 hr. After the reaction was completed, the reaction
solution was filtered by suction. The filter cake was washed with
tetrahydrofuran twice and then discarded. The filtrate was
concentrated, then dissolved in ethyl acetate (150 mL) and
distilled water (100 mL). The solution was mixed and then stood to
separate. The aqueous phase was washed again with ethyl acetate
(100 mL.times.2) twice. All the organic phases were combined, dried
over MgSO.sub.4, filtered and concentrated to give the target
compound.
Preparation 8
2-(2-T-BUTOXYCARBONYLAMINO)ETHOXY)ETHYL-4-METHYLBENZENESULFONATE
[0224] To a single-neck flask (250 mL) were added t-butyl
2-(2-hydroxyethoxy)ethylcarbamate (20.5 g) and p-toluenesulfonyl
chloride (28.575 g). Tetrahydrofuran (50 mL) was added to dissolve
the mixture. The resultant solution was cooled in an ice bath.
Sodium hydroxide (8.000 g) was dissolved in distilled water (32 g).
The solution was dropwise added in the reaction flask. The mixture
was stirred overnight. After the reaction was completed, the
reaction solution was concentrated (to remove tetrahydrofuran). To
the resultant product were added ethyl acetate (150 mL) and
distilled water (100 mL). The solution was mixed homogeneously and
stood to separate. The organic phase was washed with saturated NaCl
once, dried over MgSO.sub.4 for 30 min, filtered and concentrated
to give an oil. After standing overnight, solids were precipitated.
The solids were filtered by suction. The filter cake was eluted
with ethyl acetate twice to give the target compound.
Preparation 9
T-BUTYL 2-(2-(DIMETHYLAMINO)ETHOXY)ETHYLCARBAMATE
[0225] To a reaction flask was added dimethylamine hydrochloride
(30.922 g). Distilled water (50 mL) was added to dissolve
dimethylamine hydrochloride. The mixture was cooled in an ice bath.
To the reaction flask was added 20% sodium hydroxide aqueous
solution (76.885 g). After stirring for 20 min,
2-(2-t-butoxycarbonylamino)ethoxy)ethyl 4-methylbenzenesulfonate
(13.621 g) was dissolved in absolute ethanol (50 mL) and
tetrahydrofuran (30 mL). To the reaction flask was added the
resultant solution. The mixture reacted overnight. The reaction
flask was moved to an oil bath at 40.degree. C. The mixture was
stirred for 2.5 hr. After the reaction was completed, the organic
solvent was removed by concentration. The crude product was
extracted with ethyl acetate (150 mL.times.1) once. The pH of the
aqueous phase was adjusted with NaOH to 9. The aqueous phase was
extracted with ethyl acetate (100 mL.times.1) once. The organic
phases were combined, dried over MgSO.sub.4 for 30 min, then
filtered and concentrated to give the target compound.
Preparation 10
2-(2-(DIMETHYLAMINO)ETHOXY)ETHYLAMINE
[0226] In a reaction flask t-butyl
2-(2-(dimethylamino)ethoxy)ethylcarbamate was dissolved in
dichloromethane (70 mL). The mixture was cooled in an ice bath
under the protection of argon gas. To the reaction flask was
dropwise added trifluoroacetic acid (17 mL). The mixture reacted
overnight. After the reaction was completed, the reaction solution
was extracted with distilled water (100 mL) once. The organic phase
was discarded. The pH of the aqueous phase was adjusted with NaOH
to 13. The aqueous phase was extracted with dichloromethane (150
mL.times.3) three times. The resultant organic phase was directly
concentrated to give the target compound.
Preparation 11
T-BUTYL-DI(2-HYDROXYETHYL)CARBAMATE
[0227] To a reaction flask was added dihydroxyethylamine (31.5 g).
Tetrahydrofuran (50 mL) and distilled water (50 mL) were added to
dissolve dihydroxyethylamine. Di-t-butyl dicarbonate (85.0 g) was
dissolved in tetrahydrofuran (80 mL). The resultant solution was
dropwise added into the reaction flask in an ice bath (over 2 hr).
After the addition was completed, the mixture was stirred until the
reaction was completed (for about 1.5 hr). The reaction solution
was concentrated, then dissolved in dichloromethane (200 mL) and
distilled water (150 mL), mixed and stood to separate. The aqueous
layer was extracted with dichloromethane (100 mL.times.4) four
times. All the organic phases were combined and concentrated to
directly use in the subsequent reaction.
Preparation 12
T-BUTYL-DI(2-P-TOLUENESULFONATOETHYL)CARBAMATE
[0228] To a reaction flask were added
t-butyl-di(2-hydroxyethyl)carbamate (61.5 g) and p-toluenesulfonyl
chloride (137.2 g). Tetrahydrofuran (200 mL) was added to dissolve
t-butyl-di(2-hydroxyethyl)carbamate and p-toluenesulfonyl chloride.
The solution was cooled in an ice bath. 20% sodium hydroxide
aqueous solution (216 g) was dropwise added into the reaction flask
(over 70 min). The resultant mixture was stirred overnight in an
ice bath. After the reaction was completed, the pH of the reaction
solution was adjusted with 20% NaOH aqueous solution to 13. The
solution was stirred for 2 hr in an oil bath at 40.degree. C. The
reaction solution was concentrated. Dichloromethane (250 mL) was
added to dissolve the concentrated reaction solution. The resultant
solution was washed with distilled water (100 mL) once. The organic
phase was directly concentrated to give the crude target
compound.
Preparation 13
T-BUTYL DI(2-(DIMETHYLAMINO)ETHYL)CARBAMATE
[0229] To a reaction flask was added dimethylamine hydrochloride
(116.1 g). Distilled water (60 mL) was added to dissolve
dimethylamine hydrochloride. The solution was cooled in an ice
bath. 20% sodium hydroxide aqueous solution (284.8 g) was added
into the reaction flask (over 70 min). After the addition was
completed, the mixture was stirred for 20 min.
T-butyl-di(2-p-toluenesulfonatoethyl)carbamate (73.2 g) was
dissolved in tetrahydrofuran (200 mL). The resultant mixture was
added to the reaction flask and stirred in an oil bath at
40.degree. C. until the reaction was completed. The reaction
solution was concentrated and washed with ethyl acetate (250 mL)
once. The organic phase was re-extracted with distilled water (150
mL.times.2) twice. All the aqueous phases were combined. The pH of
the aqueous phase was adjusted with NaOH to 14. The resultant
solution was extracted with dichloromethane (200 mL.times.1, 150
mL.times.6) seven times. The seven fractions of dichloromethane
were combined, dried over anhydrous MgSO.sub.4 for 30 min, filtered
and concentrated. The resultant crude product was purified by
column chromatography (developing solvent was
CHCl.sub.3:CH.sub.3OH=15:1) to give the target compound.
Preparation 14
DI(2-(DIMETHYLAMINO)ETHYL)AMINE
[0230] To a reaction flask were added
t-butyl-di(2-(dimethylamino)ethyl)carbamate (3.000 g) and
tetrahydrofuran (20 mL). The mixture was cooled in an ice bath. To
the reaction flask was dropwise added concentrated hydrochloric
acid (9.6 mL) (over 15 min). The mixture was stirred in an ice bath
until the reaction was completed. After the reaction solution was
concentrated to remove tetrahydrofuran, the resultant solution was
washed with dichloromethane (50 mL) once. The organic phase was
discarded. The pH of the aqueous phase was adjusted with
K.sub.2CO.sub.3 to 10. The aqueous solution was extracted with
dichloromethane (50 mL.times.4) four times. The four fractions of
dichloromethane were combined, dried over anhydrous MgSO.sub.4 for
30 min, filtered and concentrated to give the target compound.
Preparation 15
2-(2-(2-(2-T-BUTOXYETHOXY)ETHOXY)ETHOXY)ETHANOL
[0231] To a reaction flask which was pre-protected with argon gas
were added tetraethylene glycol (191.2 mL) and DIEA (300 mL).
Dichloromethane (80 mL, dried via molecular sieves) was added to
the reaction flask. The mixture was dissolved with stirring at the
room temperature, and then cooled in an ice bath.
Triphenylchloromethane (205.9 g) was dissolved in dichloromethane
(400 mL, dried via molecular sieves) (over 4 hr). The resultant
mixture was added to the reaction flask and reacted overnight.
After the reaction was completed, the reaction solution was
successively washed with 5% citric acid aqueous solution (500
mL.times.4) four times and with NaCl aqueous solution (250
mL.times.1) once, dried over anhydrous MgSO.sub.4 for 30 min,
filtered and concentrated to give the target compound.
Preparation 16
2-(2-(2-(2-T-BUTOXYETHOXY)ETHOXY)ETHTHOXEY)ETHYL-4-METHYLBENZENES
ULFONATE
[0232] To a reaction flask were added
2-(2-(2-(2-t-butoxyethoxy)ethoxy)ethoxy)ethanol (347.5 g) and
p-toluenesulfonyl chloride (227.7 g). Tetrahydrofuran (200 mL) was
added to dissolve 2-(2-(2-(2-t-butoxyethoxy)ethoxy)ethoxy)ethanol
and p-toluenesulfonyl chloride. To the reaction flask was added 20%
sodium hydroxide aqueous solution (318.8 g) in an ice bath (over 2
hr). The mixture reacted overnight. After the reaction was
completed, the reaction solution was concentrated to remove
tetrahydrofuran. To the resultant crude product were added ethyl
acetate (300 mL) and distilled water (50 mL). The solution was
mixed homogeneously and stood to separate. The organic phase was
washed with saturated NaCl aqueous solution (200 mL) once, dried
over anhydrous MgSO.sub.4 for 30 min, filtered and concentrated to
give the target compound.
Preparation 17
2-(2-(2-(2-(2-T-BUTOXYETHOXY)ETHOXY)ETHOXY)ETHYL)ISOINDOL-1,3-DIONE
[0233] To a reaction flask were added
2-(2-(2-(2-t-butoxyethoxy)ethoxy)ethoxy)ethyl
4-methylbenzenesulfonate (373.7 g) and potassium phthalimide (175.7
g). DMF (350 mL, dried over molecular sieves) was added to dissolve
the mixture. The resultant solution was warmed and maintained at
65.degree. C. in an oil bath until the reaction was completed (for
about 8 hr). After the reaction solution was concentrated, ethyl
acetate (250 mL) and distilled water (200 mL) were added. The
resultant solution was mixed homogeneously and stood to separate.
The organic phase was washed with saturated NaCl aqueous solution
(150 mL) once, dried over anhydrous MgSO.sub.4 for 30 min, filtered
and concentrated to give a crude product. The crude product was
re-crystallized from anhydrous ethanol to give the target
compound.
Preparation 18
2-(2-(2-(2-T-BUTOXYETHOXY)ETHOXY))ETHOXY)ETHOXY)ETHYLAMINE
[0234] To a reaction flask was added
2-(2-(2-(2-(2-t-butoxyethoxy)ethoxy)ethoxy)ethyl)isoindol-1,3-dione
(193.515 g). Tetrahydrofuran (350 mL) was added to dissolve
2-(2-(2-(2-(2-t-butoxyethoxy)ethoxy)ethoxy)ethyl)isoindol-1,3-dione.
After 25% methylamine aqueous solution (127.410 g) was added, the
resultant solution became clear by mechanically stirring at the
room temperature. The mixture reacted overnight. After the reaction
was stirred for 30 min in an oil bath at 40.degree. C., the
reaction solution was concentrated to give white solids. These
solids were dissolved in ethyl acetate (300 mL) and distilled water
(250 mL). The resultant was mixed homogeneously and stood to
separate. The organic phase was washed with saturated NaCl aqueous
solution (200 mL) once, dried over anhydrous MgSO.sub.4 for 30 min,
filtered and concentrated to give the target compound.
Preparation 19
2-(2-(2-(-AMINOETHOXY)ETHOXY)ETHOXY)ETHANOL HYDROCHLORIDE
[0235] To a reaction flask was added concentrated hydrochloric acid
(100.5 mL). The flask was cooled in an ice bath.
2-(2-(2-(2-t-butoxyethoxy)ethoxy)ethoxy)ethylamine (123.7 g) was
dissolved in tetrahydrofuran (170 mL) (over about 2 hr). The
resultant mixture was dropwise added to the reaction flask and
reacted overnight. After the reaction was completed, the reaction
solution was concentrated to remove tetrahydrofuran. The resultant
crude product was dissolved in chloroform (150 mL) and distilled
water (100 mL). The resultant solution was mixed homogeneously and
stood to separate. The aqueous phase was directly concentrated to
give the target compound.
Preparation 20
T-BUTYL-2-(2-(2-(2-HYDROXYETHOXY)ETHOXY)ETHOXY)ETHYLCARBAMATE
[0236] To a reaction flask was added
2-(2-(2-(-aminoethoxy)ethoxy)ethoxy)ethanol hydrochloride (41.6 g).
Distilled water (80 mL) was added to dissolve
2-(2-(2-(-aminoethoxy)ethoxy)ethoxy)ethanol hydrochloride. The
solution was cooled in an ice bath. Anhydrous sodium carbonate
(38.414 g) was dissolved in distilled water (200 mL) (over 1 hr).
The mixture was dropwise added to the reaction flask. Di-t-butyl
dicarbonate (51.370 g) was dissolved in tetrahydrofuran (120 mL).
The mixture was dropwise added (over 160 min) to the reaction
flask. The resultant mixture reacted overnight. After the reaction
was completed, the reaction solution was concentrated and then
extracted with ethyl acetate (150 mL) once, extracted with
dichloromethane (150 mL) once. Two organic phases were combined,
dried over anhydrous MgSO.sub.4 for 30 min, filtered and
concentrated to give the target compound.
Preparation 21
T-BUTYL-2-(2-(2-(2-(P-METHYLPHENOXY)ETHOXY)ETHOXY)ETHOXY)ETHYL
CARBAMATE
[0237] To a reaction flask was added
t-butyl-2-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)ethylcarbamate
(14.650 g). Tetrahydrofuran (75 mL) was added to dissolve
t-butyl-2-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)ethylcarbamate. To
the reaction flask was added p-toluenesulfonyl chloride (11.439 g).
The mixture was cooled in an ice bath. To the reaction flask was
dropwise added 20% sodium hydroxide aqueous solution (18.684 g)
(over 15 min). The ice bath was removed when the addition was
completed. The resultant mixture was stirred at the room
temperature until the reaction was completed (for about 5 hr). To
the reaction flask was again added 20% sodium hydroxide aqueous
solution (8.715 g). The mixture was stirred for 2 hr in an oil bath
at 40.degree. C. The resultant solution was directly used in the
subsequent reaction.
Preparation 22
T-BUTYL-2-(2-(2-(2-(DIMETHYLAMINO)ETHOXY)ETHOXY)ETHOXY)ETHYLCARBAMATE
[0238] To a reaction flask was added dimethylamine hydrochloride
(40.766 g). Distilled water (60 mL) was added to dissolve
dimethylamine hydrochloride. The solution was cooled in an ice
bath. To the reaction flask was added 20% sodium hydroxide aqueous
solution (101.464 g). To the reaction flask was added the solution
obtained in Preparation 22. The mixture was stirred in an oil bath
at 40.degree. C. until the reaction was completed. After the
reaction solution was concentrated to remove tetrahydrofuran,
dichloromethane (200 mL) was added. The mixture was mixed
homogeneously and stood to separate. The aqueous phase was
discarded. The organic phase was added into distilled water (100
mL). The pH of the mixture was adjusted with 2 N HCl to 3. The
solution was mixed homogeneously and stood to separate. The aqueous
phase was directly concentrated to give the target compound.
Preparation 23
2-(2-(2-(2-(DIMETHYLAMINO)ETHOXY)ETHOXY)ETHOXY)ETHYLAMINE
HYDROCHLORIDE
[0239] To a reaction flask was added
t-butyl-2-(2-(2-(2-(dimethylamino)ethoxy)ethoxy)ethoxy)ethylcarbamate
(8.629 g). Tetrahydrofuran was added to dissolve
t-butyl-2-(2-(2-(2-(dimethylamino)ethoxy)ethoxy)ethoxy)ethylcarbamate.
The resultant solution was cooled in an ice bath. To the reaction
flask was added concentrated hydrochloric acid (20 mL) (over 20
min). After the reaction was completed, the reaction solution was
concentrated to remove tetrahydrofuran, and distilled water (50 mL)
and dichloromethane (50 mL) were added. The resultant mixture was
mixed homogeneously and stood to separate. The aqueous phase was
directly concentrated to give the target compound.
[0240] The target compounds in Preparation 24 to 104 were prepared
according to the preparation process in Preparation 4.
Preparation 24
4-(4-(METHYLSULFONYL)AMINO)-4-OXOBUTANOIC ACID
Preparation 25
4-(4-(PHENYLLSULFONYL)AMINO)-4-OXOBUTANOIC ACID
Preparation 26
4-(4-(MORPHOLINYLMETHYL)PHENYLAMINO)-4-OXOBUTANOIC ACID
Preparation 27
4-(4-((DIMETHYLAMINO)METHYL)PHENYLAMINO)-4-OXOBUTANOIC ACID
Preparation 28
5-(4-((DIMETHYLAMINO)METHYL)PHENYLAMINO)-5-OXOPENTANOIC ACID
Preparation 29
4-(2-(2-(DIMETHYLAMINO)ETHOXY)ETHYLAMINO)-4-OXOBUTANOIC ACID
Preparation 30
5-(2-(2-(DIMETHYLAMINO)ETHOXY)ETHYLAMINO)-5-OXOPENTANOIC ACID
Preparation 31
4-(4-(HYDROXYETHYL)PIPERAZIN-1-YL)-4-OXOBUTANOIC ACID
Preparation 32
5-(4-(HYDROXYETHYL)PIPERAZIN-1-YL)-5-OXOPENTANOIC ACID
Preparation 33
4-((MORPHOLIN-1-YL)AMINO)-4-OXOBUTANOIC ACID
Preparation 34
5-((MORPHOLIN-1-YL)AMINO)-5-OXOPENTANOIC ACID
Preparation 35
4-(3-(MORPHOLIN-1-YL)PROPYLAMINO)-4-OXOBUTANOIC ACID
Preparation 36
5-(3-(MORPHOLIN-1-YL)PROPYLAMINO)-5-OXOPENTANOIC ACID
Preparation 37
4-((4-METHYLPIPERAZIN-1-YL)AMINO)-4-OXOBUTANOIC ACID
Preparation 38
5-((4-METHYLPIPERAZIN-1-YL)AMINO)-5-OXOPENTANOIC ACID
Preparation 39
4-(2-(TETRAHYDROPYRROL-1-YL)ETHYLAMINO)-4-OXOBUTANOIC ACID
Preparation 40
5-(2-(TETRAHYDROPYRROL-1-YL)ETHYLAMINO)-5-OXOPENTANOIC ACID
Preparation 41
4-(3-(TETRAHYDROPYRROL-1-YL)PROPYLAMINO)-4-OXOBUTANOIC ACID
Preparation 42
5-(3-(TETRAHYDROPYRROL-1-YL)PROPYLAMINO)-5-OXOPENTANOIC ACID
Preparation 43
4-((6-(MORPHOLIN-1-YL)PYRIDIN-3-YL)AMINO)-4-OXOBUTANOIC ACID
Preparation 44
5-((6-(MORPHOLIN-1-YL)PYRIDIN-3-YL)AMINO)-5-OXOPENTANOIC ACID
Preparation 45
4-((PYRIDIN-4-YL)AMINO)-4-OXOBUTANOIC ACID
Preparation 46
5-((PYRIDIN-4-YL)AMINO)-5-OXOPENTANOIC ACID
Preparation 47
4-((PYRIDIN-3-YL)AMINO)-4-OXOBUTANOIC ACID
Preparation 48
4-((PYRIDIN-3-YL)AMINO)-4-OXOPENTANOIC ACID
Preparation 49
4-((PYRIDIN-4-YL)METHYLAMINO)-4-OXOBUTANOIC ACID
Preparation 50
5-((PYRIDIN-4-YL)METHYLAMINO)-5-OXOPENTANOIC ACID
Preparation 51
4-((PYRIDIN-3-YL)METHYLAMINO)-4-OXOBUTANOIC ACID
Preparation 52
5-((PYRIDIN-3-YL)METHYLAMINO)-5-OXOPENTANOIC ACID
Preparation 53
4-((PYRIDIN-2-YL)METHYLAMINO)-4-OXOBUTANOIC ACID
Preparation 54
5-((PYRIDIN-2-YL)METHYLAMINO)-5-OXOPENTANOIC ACID
Preparation 55
4-(2-(PYRIDIN-4-YL)ETHYLAMINO)-4-OXOBUTANOIC ACID
Preparation 56
5-(2-(PYRIDIN-4-YL)ETHYLAMINO)-5-OXOPENTANOIC ACID
Preparation 57
4-(2-(PYRIDIN-2-YL)ETHYLAMINO)-4-OXOBUTANOIC ACID
Preparation 58
5-(2-(PYRIDIN-2-YL)ETHYLAMINO)-5-OXOPENTANOIC ACID
Preparation 59
4-(2-(PYRIDIN-3-YL)ETHYLAMINO)-4-OXOBUTANOIC ACID
Preparation 60
5-(2-(PYRIDIN-3-YL)ETHYLAMINO)-5-OXOPENTANOIC ACID
Preparation 61
4-(2-(4-(AMINOSULFONYL)PHENYL)ETHYLAMINO)-4-OXOBUTANOIC ACID
Preparation 62
5-(2-(4-(AMINOSULFONYL)PHENYL)ETHYLAMINO)-5-OXOPENTANOIC ACID
Preparation 63
4-(4-ETHYLPIPERAZIN-1-YL)-4-OXOBUTANOIC ACID
Preparation 64
5-(4-ETHYLPIPERAZIN-1-YL)-5-OXOPENTANOIC ACID
Preparation 65
4-(PURINEAMINO)-4-OXOBUTANOIC ACID
Preparation 66
5-(PURINEAMINO)-5-OXOPENTANOIC ACID
Preparation 67
4-(4-(3-(DIMETHYLAMINO)PROPYL)PIPERAZIN-1-YL)-4-OXOBUTANOIC
ACID
Preparation 68
5-(4-(3-(DIMETHYLAMINO)PROPYL)PIPERAZIN-1-YL)-5-OXOPENTANOIC
ACID
Preparation 69
4-(3-(4-METHYLPIPERAZIN-1-YL)PROPYLAMINO)-4-OXOBUTANOIC ACID
Preparation 70
5-(3-(4-METHYLPIPERAZIN-1-YL)PROPYLAMINO)-5-OXOPENTANOIC ACID
Preparation 71
4-(2-(DIMETHYLAMINO)ETHYLAMINO)-4-OXOBUTANOIC ACID
Preparation 72
5-(2-(DIMETHYLAMINO)ETHYLAMINO)-5-OXOPENTANOIC ACID
Preparation 73
4-(DI(2-(DIMETHYLAMINO)ETHYL)AMINO)-4-OXOBUTANOIC ACID
Preparation 74
5-(DI(2-(DIMETHYLAMINO)ETHYL)AMINO)-5-OXOPENTANOIC ACID
Preparation 75
4-(2-(2-(2-(2-(DIMETHYLAMINO)ETHOXY)ETHOXY)ETHOXY)ETHYLAMINO)-4-OXOBUTANOI-
C ACID
Preparation 76
5-(2-(2-(2-(2-(DIMETHYLAMINO)ETHOXY)ETHOXY)ETHOXY)ETHYLAMINO)-5-OXOPENTANO-
IC ACID
Preparation 77
4-((PYRAZIN-2-YL)AMINO)-4-OXOBUTANOIC ACID
Preparation 78
5-((PYRAZIN-2-YL)AMINO)-5-OXOPENTANOIC ACID
Preparation 79
4-((PYRIDIN-2-YL)AMINO)-4-OXOBUTANOIC ACID
Preparation 80
5-((PYRIDIN-2-YL)AMINO)-5-OXOPENTANOIC ACID
Preparation 81
4-((PYRIMIDIN2-YL)AMINO)-4-OXOBUTANOIC ACID
Preparation 82
5-((PYRIMIDIN2-YL)AMINO)-5-OXOPENTANOIC ACID
Preparation 83
4-(2-HYDROXYETHYLAMINO)-4-OXOBUTANOIC ACID
Preparation 84
5-(2-HYDROXYETHYLAMINO)-5-OXOPENTANOIC ACID
Preparation 85
4-(2-(2-HYDROXYETHOXY)ETHYLAMINO)-4-OXOBUTANOIC ACID
Preparation 86
5-(2-(2-HYDROXYETHOXY)ETHYLAMINO)-5-OXOPENTANOIC ACID
Preparation 87
4-(DI(2-(HYDROXY)ETHYL)AMINO)-4-OXOBUTANOIC ACID
Preparation 88
5-(DI(2-(HYDROXY)ETHYL)AMINO)-5-OXOPENTANOIC ACID
Preparation 89
4-AMINO-4-OXOBUTANOIC ACID
Preparation 90
5-AMINO-5-OXOPENTANOIC ACID
Preparation 91
4-METHYLAMINO-4-OXOBUTANOIC ACID
Preparation 92
5-METHYLAMINO-5-OXOPENTANOIC ACID
Preparation 93
4-DIMETHYLAMINO-4-OXOBUTANOIC ACID
Preparation 94
5-DIMETHYLAMINO-5-OXOPENTANOIC ACID
Preparation 95
4-(MORPHOLIN-1-YL)-4-OXOBUTANOIC ACID
Preparation 96
5-(MORPHOLIN-1-YL)-5-OXOPENTANOIC ACID
Preparation 97
4-(PIPERIDIN-1-YL)-4-OXOBUTANOIC ACID
Preparation 98
4-(TETRAHYDROPYRROL-1-YL)-4-OXOBUTANOIC ACID
Preparation 99
4-(2-(MORPHOLIN-1-YL)ETHYLAMINO)-4-OXOBUTANOIC ACID
Preparation 100
4-(2-(2-(2-(2-(2-(2-(2-(2-HYDROXYETHOXY)ETHOXY)ETHOXY)ETHOXY)ETHOXY)ETHOXY-
)ETHOXY)ETHYLAMINO)-4-OXOBUTANOIC ACID
Preparation 101
4-(2-(2-(2-(2-(2-(2-(2-(2-(2-HYDROXYETHOXY)ETHOXY)ETHOXY)ETHOXY)ETHOXY)ETH-
OXY)ETHOXY)ETHOXY)ETHYLAMINO)-4-OXOBUTANOIC ACID
Preparation 102
4-(2-(2-(2-(2-(2-(2-(2-(2-(2-(2-(2-(2-HYDROXYETHOXY)ETHOXY)ETHOXY)ETHOXY)E-
THOXY)ETHOXY)ETHOXY)ETHOXY)ETHOXY)ETHOXY)ETHOXY)ETHYLAMINO)-4-OXOBUTANOIC
ACID
Preparation 103
4-(2-(2-(2-(2-(2-(2-(2-(2-(2-(METHOXY)ETHOXY)ETHOXY)ETHOXY)ETHOXY)ETHOXY)E-
THOXY)ETHOXY)ETHOXY)ETHYLAMINO)-4-OXOBUTANOIC ACID
Preparation 104
4-(2-(2-(2-(2-HYDROXYETHOXY)ETHOXY)ETHOXY)ETHYLAMINO)-4-OXOBUTANOIC
ACID
Example 1
10-((3'-(PYRROL-1-YL)-2',3',6'-TRIDEOXY-ALPHAL-L-LYXO-HEXYLPYRANYL)
OXY)-7,8,9,10-TETRAHYDRO-6,8,11-TRIHYDROXY-13-OXO-14-(4-(2-(2-(2-(2-HYDRO-
XYETHOXY)ETHOXY)ETHOXY)ETHYLAMINO)-4-OXO-BUTYRATO)-1-METH
OXY-5,12-NAPHTHALENEDIONE
[0241] To a reaction flask was added the compound (99 mg) obtained
in Preparation 1. Dichloromethane (5 mL, dried via molecular
sieves) was added to dissolve the compound. To the resultant
solution were added the compound (82 mg) obtained in Preparation
104 and DMAP (8 mg). The mixture was stirred under the protection
of argon gas. EDC-HCl (50 mg) was added. The mixture reacted
overnight at the room temperature. After the reaction was
completed, the reaction solution was concentrated and then directly
purified by thin layer chromatography (the developing solvent was
chloroform (CHCl.sub.3): methanol (CH.sub.3OH)=95: 5, 5 mL, adding
one drop of glacial acetic acid), to give the target compound. MS:
867.1 (M-1)
[0242] The compounds listed in Table 1 were prepared according to
the preparation process in Example 1, in which the dashed line in
the substituents represented the linking bond.
##STR00167##
TABLE-US-00003 TABLE 1 Examples n NR.sup.2R.sup.3 Mass Spectra (MS)
2 2 ##STR00168## MS: 868.1 (M + 1) 3 3 ##STR00169## MS: 882.3 (M +
1) 4 2 ##STR00170## MS.sup.+ 825.9 5 3 ##STR00171## MS: 840.3 (M +
1) 6 2 ##STR00172## MS.sup.- 806.2 (M - 1) MS.sup.+ 808.3 (M + 1))
7 3 ##STR00173## MS.sup.+ 822.3 (M + 1) 8 2 ##STR00174## MS.sup.+
806.1 (M + 1) 9 3 ##STR00175## MS.sup.+ 820.1 (M + 1) 10 2
##STR00176## MS.sup.-: 775.8 (M - 1) 11 3 ##STR00177## MS.sup.+:
791.2 (M + 1) 12 2 ##STR00178## MS.sup.+: 820.4 (M + 1) MS.sup.-:
818.6 (M - 1) 13 3 ##STR00179## MS.sup.+: 834.8 (M + 1) 14 2
##STR00180## MS.sup.-: 788.9 (M - 1) 15 3 ##STR00181## MS.sup.+:
805.8 (M + 1) 16 2 ##STR00182## MS.sup.+ 790.3 (M + 1) 17 3
##STR00183## MS.sup.+ 804.4 (M + 1) 18 2 ##STR00184## MS.sup.+:
804.1 (M + 1) 19 3 ##STR00185## MS.sup.+: 817.2 (M + 1) 20 2
##STR00186## MS.sup.-: 853.3 (M - 1) MS.sup.+: 855.4 (M + 1) 21 3
##STR00187## MS.sup.+: 869.3 (M + 1) 22 2 ##STR00188## MS.sup.+:
770.0 (M + 1) MS.sup.-: 767.7 (M - 1) 23 3 ##STR00189## MS.sup.+:
784.7 (M + 1) 24 2 ##STR00190## MS.sup.+: 770.1 (M + 1) 25 3
##STR00191## MS.sup.+: 784.6 (M + 1) 26 2 ##STR00192## MS.sup.+:
784.0 (M + 1) 27 3 ##STR00193## MS.sup.+: 798.2 (M + 1) 28 2
##STR00194## MS.sup.+: 783.9 (M + 1) MS.sup.-: 781.8 (M - 1) 29 3
##STR00195## MS.sup.+: 798.9 (M + 1) 30 2 ##STR00196## MS.sup.+:
784.1 (M + 1) 31 3 ##STR00197## MS.sup.+: 798.1 (M + 1) 32 2
##STR00198## MS.sup.+: 798.0 (M + 1) 33 3 ##STR00199## MS.sup.+:
812.1 (M + 1) 34 2 ##STR00200## MS.sup.+: 798.0 (M + 1) 35 3
##STR00201## MS.sup.+: 812.3 (M + 1) 36 2 ##STR00202## MS.sup.+:
798.0 (M + 1) 37 3 ##STR00203## MS.sup.+: 812.1 (M + 1) 38 2
##STR00204## MS.sup.-: 873.7 (M - 1) 39 3 ##STR00205## MS.sup.-:
888.1 (M - 1) 40 2 ##STR00206## MS.sup.+: 790.1 (M + 1) 41 3
##STR00207## MS.sup.+: 804.3 (M + 1) 42 2 ##STR00208## MS.sup.+:
811.0 (M + 1) MS.sup.-: 809.2 (M - 1) 43 3 ##STR00209## MS.sup.+:
825.4 (M + 1) 44 2 ##STR00210## MS.sup.+: 847.1 (M + 1) MS.sup.-:
845.0 (M - 1) 45 3 ##STR00211## MS.sup.+: 860.3 (M + 1) 46 2
##STR00212## MS.sup.+: 833.1 (M + 1) 47 3 ##STR00213## MS.sup.+:
847.2 (M + 1) 48 2 ##STR00214## MS.sup.+: 764.1 (M + 1) 49 3
##STR00215## MS.sup.+: 778.1 (M + 1) 50 2 ##STR00216## MS.sup.+:
835.2 (M + 1) 51 3 ##STR00217## MS.sup.+: 849.2 (M + 1) 52 2
##STR00218## MS.sup.+ 896.1 (M + 1) 53 3 ##STR00219## MS.sup.+
909.8 (M + 1) 54 2 ##STR00220## MS.sup.+ 771.2 (M + 1) 55 3
##STR00221## MS.sup.+ 784.1 (M + 1) 56 2 ##STR00222## MS.sup.+
770.1 (M + 1) 57 3 ##STR00223## MS.sup.+ 784.1 (M + 1) 58 2
##STR00224## MS.sup.+ 770.1 (M + 1) 59 3 ##STR00225## MS.sup.+
784.1 (M + 1) 60 2 ##STR00226## MS.sup.-: 735.2 (M - 1) MS.sup.+:
759.3 (M + Na.sup.+) 61 3 ##STR00227## MS.sup.-: 750.7 (M - 1) 62 2
##STR00228## MS.sup.+: 803.3 (M + Na.sup.+) 63 3 ##STR00229##
MS.sup.-: 793.6 (M - 1) 64 2 ##STR00230## MS.sup.+: 803.2 (M +
Na.sup.+) MS.sup.-: 799.2 (M - 1) 65 3 ##STR00231## MS.sup.+: 795.2
(M + 1) 66 2 NH.sub.2 MS.sup.+: 693.1 (M + 1) 67 3 NH.sub.2
MS.sup.+: 707.2 (M + 1) 68 2 NHCH.sub.3 MS.sup.-: 705.6 (M - 1) 69
3 NHCH.sub.3 MS.sup.+: 721.1 (M + 1) 70 2 N(CH.sub.3).sub.2
MS.sup.+: 721.1 (M + 1) 71 3 N(CH.sub.3).sub.2 MS.sup.+: 735.6 (M +
1) 72 2 ##STR00232## MS.sup.+: 763.2 (M + 1) 73 3 ##STR00233##
MS.sup.+: 777.1 (M + 1) 74 2 ##STR00234## MS.sup.+: 761.3 (M + 1)
75 2 ##STR00235## MS.sup.+: 746.9 (M + 1) 88 2 ##STR00236##
MS.sup.+: 806.2 (M + 1) 90 2 ##STR00237## 91 2 ##STR00238## 92 2
##STR00239## 93 2 ##STR00240## 94 2 ##STR00241## MS.sup.+: 771 (M +
1) 95 2 ##STR00242## MS.sup.+: 833 (M + 1) 97 3 ##STR00243## 98 2
##STR00244##
[0243] The .sup.1H-NMR spectra data of the compound in Example 28
were:
[0244] .delta.=1.145 ppm (d, 3H), .delta.=1.664 ppm (m, 1H),
.delta.=2.110 ppm (m, 1H),
[0245] .delta.=2.336 ppm (d, 1H), .delta.=2.415 ppm (m, 1H),
.delta.=2.505 ppm (m, 2H),
[0246] .delta.=2.675 ppm (t, 2H), .delta.=2.917 ppm (d, 1H),
.delta.=3.008 ppm (d, 1H),
[0247] .delta.=3.546 ppm (s, 1H), .delta.=3.943 ppm (s, 4H),
.delta.=4.303 ppm (m, 5H),
[0248] .delta.=4.982 ppm (s, 1H), .delta.=5.189 ppm (d, 1H),
.delta.=5.253 ppm (d, 1H),
[0249] .delta.=5.349 ppm (s, 1H), .delta.=5.920 ppm (s, 2H),
.delta.=6.780 ppm (s, 2H),
[0250] .delta.=7.346 ppm (m, 1H), .delta.=7.609 ppm (d, 1H),
.delta.=7.640 ppm (d, 1H),
[0251] .delta.=7.856 ppm (m, 2H), .delta.=8.471 ppm (m, 3H),
.delta.=13.219 ppm (s, 1H),
[0252] .delta.=13.982 ppm (s, 1H)
[0253] The .sup.1H-NMR spectra data of the compound in Example 88
were:
[0254] .delta.=1.149 ppm (d, 3H), .delta.=2.646 ppm (d, 2H),
.delta.=2.409 ppm (t, 2H),
[0255] .delta.=1.375-2.445 ppm (m, 2H), .delta.=2.795-2.936 ppm
(dd, 2H),
[0256] .delta.=2.702 ppm (s, 4H), .delta.=4.294 ppm (m, 1H),
.delta.=3.869 ppm (s, 3H),
[0257] .delta.=2.632 ppm (d, 2H), .delta.=3.666 ppm (t, 4H),
[0258] .delta.=5.162-5.236 ppm (dd, 2H), .delta.=4.282 ppm (m,
1H),
[0259] .delta.=4.897 ppm (s, 1H), .delta.=3.533 ppm (s, 1H),
.delta.=5.314 ppm (s, 1H),
[0260] .delta.=5.899 ppm (t, 2H), .delta.=7.729 ppm (d, 1H),
.delta.=6.755 ppm (t, 2H),
[0261] .delta.=7.492 ppm (d, 1H), .delta.=7.772 ppm (t, 1H),
.delta.=8.070 ppm (t, 1H),
[0262] .delta.=13.097 ppm (bs, 1H), .delta.=13.871 ppm (s, 1H)
Example 76
10-((3'-(PYRROL-1-YL)-2',3',6'-TRIDEOXY-ALPHAL-L-LYXO-HEXYLPYRANYL)
OXY)-7,8,9,10-TETRAHYDRO-6,
8,11-TRIHYDROXY-13-OXO-14-(4-(2-(2-(DIMETH
YLAMINO)ETHOXY)ETHYLAMINO)-4-OXO-BUTYRATO)-1-METHOXY-5,12-NAPHTHALENEDION-
E ACETATE
[0263] The target compound (10 mg) obtained in Example 6 was
dissolved in chloroform (10 mL) with stirring. To a reaction flask
was added glacial acetic acid (1 mg) dissolved in chloroform (10
mL). The mixture was continuously stirred for 10 min, and
rotary-evaporated to remove the solvent, thereby giving the target
compound.
[0264] The solubility of the compound in water was more than 9
mg/mL.
[0265] The target compounds in Examples 77-87 and 89 were prepared
according to the preparation process in Example 76.
Example 77
10-((3'-(PYRROL-1-YL)-2',3',6'-TRIDEOXY-ALPHAL-L-LYXO-HEXYLPYRANYL)
OXY)-7,8,9,10-TETRAHYDRO-6,8,11-TRIHYDROXY-13-OXO-14-(4-((4-(2-HYDROXY)ET-
HYL)PIPERAZIN-1-YL)-4-OXO-BUTYRATO)-1-METHOXY-5,12-NAPHTHALENEDIONE
ACETATE
[0266] The solubility of the compound in water was more than 10.6
mg/ml.
Example 78
10-((3'-(PYRROL-1-YL)-2',3',6'-TRIDEOXY-ALPHAL-L-LYXO-HEXYLPYRANYL)
OXY)-7,8,9,10-TETRAHYDRO-6,8,11-TRIHYDROXY-13-OXO-14-(4-((3-(MORPHOLIN-1--
YL)PROPYL)AMINO)-4-OXO-BUTYRATO)-1-METHOXY-5,12-NAPHTHALENE DIONE
ACETATE
[0267] The solubility of the compound in water was more than 22
mg/ml.
Example 79
10-((3'-(PYRROL-1-YL)-2',3',6'-TRIDEOXY-ALPHAL-L-LYXO-HEXYLPYRANYL)
OXY)-7,8,9,10-TETRAHYDRO-6,8,11-TRIHYDROXY-13-OXO-14-(4-((4-METHYL)PIPERA-
ZIN-1-YL)AMINO)-4-OXO-BUTYRATO)-1-METHOXY-5,12-NAPHTHALENE DINOE
ACETATE
[0268] The solubility of the compound in water was more than 5
mg/ml.
Example 80
10-((3'-(PYRROL-1-YL)-2',3',6'-TRIDEOXY-ALPHAL-L-LYXO-HEXYLPYRANYL)
OXY)-7, 8,9,10-TETRAHYDRO-6,
8,11-TRIHYDROXY-13-OXO-14-(4-(4-ETHYLPIPERAZIN-1-YL)-4-OXO-BUTYRATO)-1-ME-
THOXY-5,12-NAPHTHALENEDIONE ACETATE
[0269] The solubility of the compound in water was more than 11
mg/ml.
Example 81
10-((3'-(PYRROL-1-YL)-2',3',6'-TRIDEOXY-ALPHAL-L-LYXO-HEXYLPYRANYL)
OXY)-7,8,9,10-TETRAHYDRO-6,8,11-TRIHYDROXY-13-OXO-14-(4-((3-(4-METHYL
PIPERAZIN-1-YL)PROPYL)AMINO)-4-OXO-BUTYRATO)-1-METHOXY-5,12-NAPHTHALENEDI-
ONE ACETATE
[0270] The solubility of the compound in water was more than 7
mg/ml.
Example 82
10-((3'-(PYRROL-1-YL)-2',3',6'-TRIDEOXY-ALPHAL-L-LYXO-HEXYLPYRANYL)
OXY)-7,8,9,10-TETRAHYDRO-6,
8,11-TRIHYDROXY-13-OXO-14-(4-((PYRIDIN-4-YL)METHYL)AMINO)-4-OXO-BUTYRATO)-
-1-METHOXY-5,12-NAPHTHALENEDIONE ACETATE
[0271] The solubility of the compound in water was more than 6
mg/ml.
Example 83
10-((3'-(PYRROL-1-YL)-2',3',6'-TRIDEOXY-ALPHAL-L-LYXO-HEXYLPYRANYL)
OXY)-7,8,9,10-TETRAHYDRO-6,8,11-TRIHYDROXY-13-OXO-14-(4-((PYRIDIN-3-YL)ME-
THYL)AMINO)-4-OXO-BUTYRATO)-1-METHOXY-5,12-NAPHTHALENEDIONE
ACETATE
[0272] The solubility of the compound in water was more than 1
mg/ml.
Example 84
10-((3'-(PYRROL-1-YL)-2',3',6'-TRIDEOXY-ALPHAL-L-LYXO-HEXYLPYRANYL)
OXY)-7,
8,9,10-TETRAHYDRO-6,8,11-TRIHYDROXY-13-OXO-14-(4-(2-(PYRIDIN-2-YL-
)ETHYL)AMINO)-4-OXO-BUTYRATO)-1-METHOXY-5,12-NAPHTHALENEDIONE
ACETATE
[0273] The solubility of the compound in water was more than 12
mg/ml.
Example 85
10-((3'-(PYRROL-1-YL)-2',3',6'-TRIDEOXY-ALPHAL-L-LYXO-HEXYLPYRANYL)
OXY)-7,
8,9,10-TETRAHYDRO-6,8,11-TRIHYDROXY-13-OXO-14-(4-(2-(PYRIDIN-3-YL-
)ETHYL)AMINO)-4-OXO-BUTYRATO)-1-METHOXY-5,12-NAPHTHALENEDIONE
ACETATE
[0274] The solubility of the compound in water was more than 11
mg/ml.
Example 86
10-((3'-(PYRROL-1-YL)-2',3',6'-TRIDEOXY-ALPHAL-L-LYXO-HEXYLPYRANYL)
OXY)-7,8,9,10-TETRAHYDRO-6,8,11-TRIHYDROXY-13-OXO-14-(4-(2-(PYRIDIN-4-YL)-
ETHYL)AMINO)-4-OXO-BUTYRATO)-1-METHOXY-5,12-NAPHTHALENEDIONE
ACETATE
[0275] The solubility of the compound in water was more than 3
mg/ml.
Example 87
10-((3'-(PYRROL-1-YL)-2',3',6'-TRIDEOXY-ALPHAL-L-LYXO-HEXYLPYRANYL)
OXY)-7,8,9,10-TETRAHYDRO-6,
8,11-TRIHYDROXY-13-OXO-14-(4-(4-(3-(DIMETH
YLAMINO)PROPYL)PIPERAZIN-1-YL)-4-OXO-BUTYRATO)-1-METHOXY-5,12-NAPHTHALENE-
DIONE ACETATE
[0276] The solubility of the compound in water was more than 16
mg/ml.
Example 89
10-((3'-(PYRROL-1-YL)-2',3',6'-TRIDEOXY-ALPHAL-L-LYXO-HEXYLPYRANYL)
OXY)-7,8,9,10-TETRAHYDRO-6,8,11-TRIHYDROXY-13-OXO-14-(4-((2-(MORPHOLIN-1--
YL)ETHYL)AMINO)-4-OXO-BUTYRATO)-1-METHOXY-5,12-NAPHTHALENE DIONE
ACETATE
Example 96
10-((3'-(PYRROL-1-YL)-2',3',6'-TRIDEOXY-ALPHAL-L-LYXO-HEXYLPYRANYL)
OXY)-7,8,9,10-TETRAHYDRO-6,8,11-TRIHYDROXY-13-OXO-14-(4-((2-(MORPHOLIN-1--
YL)ETHYL)AMINO)-4-OXO-BUTYRATO)-1-METHOXY-5,12-NAPHTHALENE DIONE
PHOSPHATE
[0277] The solubility of the compound in water was more than 26
mg/ml.
[0278] The compound in Example 96 was detected by HPLC after being
sealed and stored for 9 months under refrigeration conditions. No
obvious degraded product was observed, which indicates good
stability of the compound.
Example 99
10-((3'-(PYRROL-1-YL)-2',3',6'-TRIDEOXY-ALPHAL-L-LYXO-HEXYLPYRANYL)
OXY)-7,8,9,10-TETRAHYDRO-6,8,11-TRIHYDROXY-13-OXO-14-(4-((2-(MORPHOLIN-1--
YL)ETHYL)AMINO)-4-OXO-BUTYRATO)-1-METHOXY-5,12-NAPHTHALENE DIONE
HYDROCHLORIDE
Biological Activity Examples
Example 1
SK-OV-3 Cell Growth Inhibition Test (MTT Assay)
I. Assay Materials
[0279] Cell strains: SK-OV-3 (human ovarian cancer cell strains);
MTT; antitumor compounds; DMSO.
II. Reagents and Consumable Materials
[0280] Culture medium: 90% McCoy's 5A+10% FBS; Pancreatin (0.25%
(w/v) solution was formulated with PBS, 0.53 mM of EDTA was added
in the formulation); PBS; 96-well culture plate.
III. Assay Process
[0281] 1. A plate (10 cm) of cells in logarithmic growth phase that
were normally cultured was collected;
[0282] 2. The culture solution was sucked out. The plate was washed
with 5 mL of PBS once or twice;
[0283] 3. PBS was sucked out. 1.5 mL of 0.25% pancreatin was added
to infiltrate the cells;
[0284] 4. The pancreatin was sucked out. The culture plate was
placed in an incubator. Digestion was carried out for about 5 min
at 37.degree. C.;
[0285] 5. 4.5 mL of complete culture solution was added to the
culture plate to stop the digestion. The cells were carefully
scoured with micropipette (1 ml) to give a uniform cell suspension.
The suspension was implanted into a 96-well cell culture disc by
4000 cells/100 .mu.L per well. The culture plate was incubated
overnight under 5% CO.sub.2 at 37.degree. C. In day 2, 100 .mu.L of
culture solution comprising a compound was added in each well. The
plate was further incubated for 69 h under 5% CO.sub.2 at
37.degree. C.;
[0286] 6. The culture solution was sucked out;
[0287] 7. 100 .mu.L of serum-free culture solution containing 0.5
mg/mL MTT was added in each well. The plate was incubated for 3
h;
[0288] 8. The culture solution was carefully sucked out;
[0289] 9. 100 .mu.L of DMSO was added in each well and vibrated to
dissolve;
[0290] 10. OD values were determined at 490 nM.
IV. Results and Treatments
[0291] 1. Calculation of Relative Inhibition Ratio
The inhibition ratio of a compound on cell
growth=(PC-n)/(PC-NC).times.100%
[0292] wherein:
[0293] PC: OD values of cells after normal growth in control wells
without a compound;
[0294] n: OD values of cells after growth in test wells with a
compound;
[0295] NC: Background OD values of blank wells without a compound
and cells.
TABLE-US-00004 TABLE 2 Growth Inhibition Activity at 667 nM of Some
Compounds in Examples on SK-OV-3 Cells Inhi- bition Inhibition
Inhibition Ratio Compounds Ratio (%) Compounds Ratio (%) Compounds
(%) Example 1 57.3 Example 3 62.5 Example 8 60 Example 10 71.5
Example 12 48.8 Example 14 53.6 Example 18 47.4 Example 20 62.6
Example 22 60.3 Example 24 61.4 Example 26 67.6 Example 28 66.2
Example 30 59.9 Example 32 59.7 Example 34 59.4 Example 35 57.8
Example 38 67.6 Example 40 47.9 Example 44 52.3 Example 60 58.9
Example 62 63.6 Example 64 65.9 Example 66 66.5 Example 76 46.4
Example 77 60.4 Example 81 56.4 Example 94 71.4 Example 95 66.3
Example 2
Bxpc-3 Cell Growth Inhibition Test (Mtt Assay)
I. Assay Materials:
[0296] Cell strains: BxPC-3 (human pancreatic cancer cell strains);
MTT; antitumor compounds; DMSO.
II. Reagents and Consumable Materials
[0297] Culture medium: 90% RPMI-1640+10% FBS; Pancreatin (0.25%
(w/v) solution was formulated with PBS, 0.53 mM of EDTA was added
in the formulation); PBS; 96-well culture plate.
III. Assay Process:
[0298] 1. A plate (10 cm) of cells in logarithmic growth phase that
were normally cultured was collected;
[0299] 2. The culture solution was sucked out. The plate was washed
with 5 mL of PBS once or twice;
[0300] 3. PBS was sucked out. 1.5 mL of 0.25% pancreatin was added
to infiltrate the cells;
[0301] 4. The culture plate was placed in an incubator. Digestion
was carried out for about 8 min at 37.degree. C.;
[0302] 5. 4.5 mL of complete culture solution was added to the
culture plate to stop the digestion. The cells were carefully
scoured with micropipette (1 ml) to give a uniform cell suspension.
The suspension was implanted into a 96-well cell culture disc by
5000 cells/100 .mu.L per well. The culture plate was incubated
overnight under 5% CO.sub.2 at 37.degree. C. In day 2, 100 .mu.L of
culture solution comprising a compound was added in each well. The
plate was further incubated for 70 h under 5% CO.sub.2 at
37.degree. C.;
[0303] 6. The culture solution was sucked out;
[0304] 7. 100 .mu.L of serum-free culture solution containing 0.5
mg/mL MTT was added in each well. The plate was incubated for 3
h;
[0305] 8. The culture solution was carefully sucked out;
[0306] 9. 100 .mu.L of DMSO was added into each well and vibrated
to dissolve;
[0307] 10. OD values were determined at 490 nM.
IV. Results and Treatments
[0308] 1. Calculation of Relative Inhibition Ratio
The inhibition ratio of a compound on cell
growth=(PC-n)/(PC-NC).times.100%
[0309] wherein:
[0310] PC: OD values of cells after normal growth in control wells
without a compound;
[0311] n: OD values of cells after growth in test wells with a
compound;
[0312] NC: Background OD values of blank wells without a compound
and cells.
TABLE-US-00005 TABLE 3 Growth Inhibition Activity at 2000 nM of
Some Compounds in Examples on BxPc-3 Cells Inhi- bition Inhibition
Inhibition Ratio Compounds Ratio (%) Compounds Ratio (%) Compounds
(%) Example 1 82.5 Example 3 81.2 Example 8 82.6 Example 10 85
Example 12 74.9 Example 14 81.8 Example 18 70.7 Example 20 84.1
Example 22 82 Example 24 84.7 Example 26 82.7 Example 28 84.7
Example 30 85.2 Example 32 83.4 Example 34 84 Example 35 81.3
Example 38 82.4 Example 40 76.5 Example 44 78.5 Example 60 83.4
Example 62 82.5 Example 64 83.7 Example 66 83.1 Example 76 77.1
Example 77 83.3 Example 81 80.7 Example 94 84.9 Example 95 80.6
Example 3
NCI-H446 Cell Growth Inhibition Test (MTT Assay)
I. Assay Materials
[0313] Cell strains: NCI-H446 (human small cell lung cancer cell
strains); MTT; antitumor compounds; DMSO.
II. Reagents and Consumable Materials
[0314] Culture medium: 90% RPMI-1640+10% FBS; Pancreatin (0.25%
(w/v) solution was formulated with PBS, 0.53 mM of EDTA was added
in the formulation); PBS; 96-well culture plate.
III. Assay Process
[0315] 1. A plate (10 cm) of cells in logarithmic growth phase that
were normally cultured was collected;
[0316] 2. The culture solution was sucked out. The plate was washed
with 5 mL of PBS once or twice;
[0317] 3. PBS was sucked out. 1.5 mL of 0.25% pancreatin was added
to infiltrate the cells;
[0318] 4. The culture plate was placed in an incubator. Digestion
was carried out for about 3 min at 37.degree. C.;
[0319] 5. 3 mL of complete culture solution was added to the
culture plate to stop the digestion. The cells were carefully
scoured with micropipette (1 ml) to give a uniform cell suspension.
The suspension was implanted into a 96-well cell culture disc by
4000 cells/100 .mu.L per well. The culture plate was incubated
overnight under 5% CO.sub.2 at 37.degree. C. In day 2, 100 .mu.L of
culture solution comprising a compound was added in each well. The
plate was further incubated for 70 h under 5% CO.sub.2 at
37.degree. C.;
[0320] 6. The culture solution was sucked out;
[0321] 7. 100 .mu.L of serum-free culture solution containing 0.5
mg/mL MTT was added in each well. The plate was incubated for 3
h;
[0322] 8. The culture solution was carefully sucked out;
[0323] 9. 100 .mu.L of DMSO was added into each well and vibrated
to dissolve;
[0324] 10. OD values were determined at 490 nM.
IV. Results and Treatments
[0325] 1. Calculation of Relative Inhibition Ratio
The inhibition ratio of a compound on cell
growth=(PC-n)/(PC-NC).times.100%
[0326] wherein:
[0327] PC: OD values of cells after normal growth in control wells
without a compound;
[0328] n: OD values of cells after growth in test wells with a
compound;
[0329] NC: Background OD values of blank wells without a compound
and cells.
TABLE-US-00006 TABLE 4 Growth Inhibition Activity at 222 nM of Some
Compounds in Examples on NCI-H446 Cells Inhi- bition Inhibition
Inhibition Ratio Compounds Ratio (%) Compounds Ratio (%) Compounds
(%) Example 1 43.2 Example 3 60.8 Example 8 59.5 Example 10 61.1
Example 12 54.6 Example 20 68 Example 22 62.1 Example 24 61.8
Example 26 66.9 Example 28 65.8 Example 30 61 Example 32 59.3
Example 34 60.7 Example 35 57.2 Example 38 62.6 Example 40 49
Example 44 46.3 Example 60 52.5 Example 62 64.6 Example 64 51.6
Example 66 56.2 Example 76 46 Example 77 59.1 Example 81 56.8
Example 94 48.4
Example 4
MDA-Mb-453 Cell Growth Inhibition Test (SRB Assay)
I. Assay Materials
[0330] Cell strains: MDA-MB-453 (human breast cancer cell strains);
SRB: 0.4% (w/v) working solution was formulated with 1% glacial
acetic acid, reserved at 4.degree. C.; antitumor compounds;
DMSO.
II. Reagents and Consumable Materials
[0331] Culture medium: 90% L15+10% FBS; pancreatin (0.25% (w/v)
solution was formulated with PBS, 0.53 mM of EDTA was added in the
formulation); PBS; 96-well culture plate.
III. Assay Process
[0332] 1. A plate (10 cm) of cells in logarithmic growth phase that
were normally cultured was collected;
[0333] 2. The culture solution was sucked out. The plate was washed
with 5 mL of PBS once or twice;
[0334] 3. 1.5 mL of 0.25% pancreatin was added to the plate to
infiltrate the cells;
[0335] 4. The pancreatin was sucked out. The culture plate was
placed in an incubator. Digestion was carried out for about 3 min
at 37.degree. C.;
[0336] 5. 4 mL of complete culture solution was added to the
culture plate to stop the digestion. The cells were carefully
scoured with micropipette (1 ml) to give a uniform single cell
suspension. The suspension was implanted into a 96-well cell
culture plate by 7000 cells/100 .mu.L per well. The culture plate
was incubated overnight under 5% CO.sub.2 at 37.degree. C. In day
2, 100 .mu.L of culture solution comprising a compound was added in
each well. The plate was further incubated for 69.5 h under 5%
CO.sub.2 at 37.degree. C.;
[0337] 6. The culture solution was sucked out. 100 uL of TCA fixed
cells which were diluted to 10% were added to each well. The plate
was kept in a refrigerator for 1 h at 4.degree. C. 7. TCA
stationary liquid was sucked out. Each well was washed with 150
.mu.L of ddH.sub.2O five times;
[0338] 8. After the stationary liquid was cleansed, the plate was
dried in the air at the room temperature;
[0339] 9. 60 .mu.L of SRB staining solution was added in each well.
The well was stained for 15 min at the room temperature;
[0340] 10. The SRB staining solution was sucked out. Each well was
washed with 150 .mu.L of 1% glacial acetic acid five times;
[0341] 11. After the SRB staining solution was cleansed, the plate
was dried in the air at the room temperature;
[0342] 12. 100 .mu.L of 10 mM Tris was added in each well. The
plate was vibrated to dissolve out SRB;
[0343] 13. OD values were determined at 570 nM.
IV. Results and Treatments
[0344] 1. Calculation of Relative Inhibition Ratio
The inhibition ratio of a compound on cell
growth=(PC-n)/(PC-NC).times.100%
[0345] wherein:
[0346] PC: OD values of cells after normal growth in control wells
without a compound;
[0347] n: OD values of cells after growth in test wells with a
compound;
[0348] NC: Background OD values of blank wells without a compound
and cells.
TABLE-US-00007 TABLE 5 Growth Inhibition Activity at 2000 nM of
Some Compounds in Examples on MDA-MB-453 Cells Inhi- bition
Inhibition Inhibition Ratio Compounds Ratio (%) Compounds Ratio (%)
Compounds (%) Example 1 77.7 Example 3 73.9 Example 8 75.3 Example
10 78.5 Example 12 73.7 Example 14 74.4 Example 20 82.4 Example 22
71.1 Example 24 79.8 Example 26 79.9 Example 28 81.9 Example 30
79.9 Example 32 78.3 Example 34 80.5 Example 35 77.1 Example 38
75.8 Example 40 73 Example 44 68.5 Example 60 76.3 Example 62 83.7
Example 64 73.5 Example 66 73.9 Example 76 62.7 Example 77 79.7
Example 81 72 Example 94 75.8 Example 95 68.2
Example 5
22Rv1 Cell Growth Inhibition Test (SRB Assay)
I. Assay Materials
[0349] Cell strains: 22Rv1 (human prostate cancer cell strains);
SRB: 0.4% (w/v) working solution was formulated with 1% glacial
acetic acid, reserved at 4.degree. C.; antitumor compounds;
DMSO.
II. Reagents and Consumable Materials
[0350] Culture medium: 90% RPMI-1640+10% FBS; Pancreatin (0.25%
(w/v) solution was formulated with PBS, 0.53 mM of EDTA was added
in the formulation); PBS; 96-well culture plate.
III. Assay Process
[0351] 1. A plate (10 cm) of cells in logarithmic growth phase that
were normally cultured was collected;
[0352] 2. The culture solution was sucked out. The plate was washed
with 5 mL of PBS once or twice;
[0353] 3. 1.5 mL of 0.25% pancreatin was added to the plate to
infiltrate the cells;
[0354] 4. The pancreatin was sucked out. The culture plate was
placed in an incubator. Digestion was carried out for about 3 min
at 37.degree. C.;
[0355] 5. 4 mL of complete culture solution was added to the
culture plate to stop the digestion. The cells were carefully
scoured with micropipette (1 ml) to give a uniform single cell
suspension. The suspension was implanted into a 96-well cell
culture plate by 7000 cells/100 .mu.L per well. The culture plate
was incubated overnight under 5% CO.sub.2 at 37.degree. C. In day
2, 100 .mu.L of culture solution comprising a compound was added in
each well. The plate was further incubated for 73 h under 5%
CO.sub.2 at 37.degree. C.;
[0356] 6. The culture solution was sucked out. 100 uL of TCA fixed
cells which were diluted to 10% were added to each well. The plate
was kept in a refrigerator for 1 h at 4.degree. C.
[0357] 7. TCA stationary liquid was sucked out. Each well was
washed with 150 .mu.L of ddH.sub.2O five times;
[0358] 8. After the stationary liquid was cleansed, the plate was
dried in the air at the room temperature;
[0359] 9. 60 .mu.L of SRB staining solution was added in each well.
The well was stained for 15 min at the room temperature;
[0360] 10. The SRB staining solution was sucked out. Each well was
washed with 150 .mu.L of 1% glacial acetic acid five times;
[0361] 11. After the SRB staining solution was cleansed, the plate
was dried in the air at the room temperature;
[0362] 12. 100 .mu.L of 10 mM Tris was added in each well. The
plate was vibrated to dissolve out SRB;
[0363] 13. OD values were determined at 570 nM.
IV. Results and Treatments
[0364] 1. Calculation of Relative Inhibition Ratio
The inhibition ratio of a compound on cell
growth=(PC-n)/(PC-NC).times.100%
[0365] wherein:
[0366] PC: OD values of cells after normal growth in control wells
without a compound;
[0367] n: OD values of cells after growth in test wells with a
compound;
[0368] NC: Background OD values of blank wells without a compound
and cells.
TABLE-US-00008 TABLE 6 Growth Inhibition Activity at 222 nM of Some
Compounds in Examples on 22Rv1 Cells Inhi- bition Inhibition
Inhibition Ratio Compounds Ratio (%) Compounds Ratio (%) Compounds
(%) Example 1 52.2 Example 3 61.9 Example 8 56.8 Example 10 62.7
Example 12 53.3 Example 14 52.1 Example 20 60.9 Example 22 56.3
Example 24 63.6 Example 26 60.8 Example 28 63.1 Example 30 62.3
Example 32 61.7 Example 34 63.3 Example 35 58.5 Example 38 56.3
Example 40 50.5 Example 44 48.5 Example 60 52.9 Example 62 57
Example 64 54.8 Example 66 59 Example 76 49.2 Example 77 57.9
Example 81 56.5 Example 94 60.7 Example 95 51.3
Example 6
A375 Cell Growth Inhibition Test (MTT Assay)
I. Assay Materials
[0369] Cell strains: A375 (human cutaneous melanoma cell strains);
MTT: nitroblue tetrazolium; antitumor compounds; compound A:
3'-pyrrolyldoxorubicin-14-oxo-succinic acid monoester; DMSO.
II. Reagents and Consumable Materials
[0370] Culture medium: 90% DMEM+10% FBS (fetal calf serum);
Pancreatin (0.25% (w/v) solution was formulated with PBS, 0.53 mM
of EDTA was added in the formulation); PBS; 96-well culture
plate.
III. Assay Process
[0371] 1. A plate (10 cm) of cells in logarithmic growth phase that
were normally cultured was collected;
[0372] 2. The culture solution was sucked out. The plate was washed
with 5 mL of PBS once or twice;
[0373] 3. PBS was sucked out. 1.5 mL of 0.25% pancreatin was added
to the plate to infiltrate the cells for 1 min;
[0374] 4. The culture plate was placed in an incubator. Digestion
was carried out for about 5 min at 37.degree. C.;
[0375] 5. 3 mL of complete culture solution was added to the
culture plate to stop the digestion. The cells were carefully
scoured with micropipette (1 ml) to give a uniform cell suspension.
The suspension was implanted into a 96-well cell culture disc by
3000 cells/100 .mu.L per well. The culture plate was incubated
overnight under 5% CO.sub.2 at 37.degree. C. In day 2, 100 .mu.L of
culture solution comprising a compound was added in each well. The
plate was further incubated for 72 h under 5% CO.sub.2 at
37.degree. C.;
[0376] 6. The culture solution was sucked out;
[0377] 7. 100 .mu.L of serum-free culture solution containing 0.5
mg/mL MTT was added in each well. The plate was incubated for 3
h;
[0378] 8. The culture solution was carefully sucked out;
[0379] 9. 100 .mu.L of DMSO was added in each well and vibrated to
dissolve;
[0380] 10. OD values were determined at 490 nM.
IV. Results and Treatments
[0381] 1. Calculation of Relative Inhibition Ratio
The inhibition ratio of a compound on cell
growth=(PC-n)/(PC-NC).times.100%
[0382] wherein:
[0383] PC: OD values of cells after normal growth in control wells
without a compound;
[0384] n: OD values of cells after growth in test wells with a
compound;
[0385] NC: Background OD values of blank wells without a compound
and cells.
TABLE-US-00009 TABLE 7 Growth Inhibition Activity of Some Compounds
in Examples on A375 Cells Inhibition Inhibition Inhibition Activity
Com- Activity Com- Activity Compounds (IC.sub.50/nM) pounds
(IC.sub.50/nM) pounds (IC.sub.50/nM) Compound 207 Example 67
Example 48 A 6 89 Example 81 67 Example 28 Example 35 97 99
Example 7
A431 Cell Growth Inhibition Test (MTT Assay)
I. Assay Materials
[0386] Cell strains: A431 (human epidermal carcinoma cell strains);
MTT: nitroblue tetrazolium; antitumor compounds; compound A:
3'-pyrrolyldoxorubicin-14-oxo-succinic acid monoester; DMSO.
II. Reagents and Consumable Materials
[0387] Culture medium: 45% DMEM+45% F12+10% FBS (fetal calf serum);
Pancreatin (0.25% (w/v) solution was formulated with PBS, 0.53 mM
of EDTA was added in the formulation); PBS; 96-well culture
plate.
III. Assay Process
[0388] 1. A plate (10 cm) of cells in logarithmic growth phase that
were normally cultured was collected;
[0389] 2. The culture solution was sucked out. The plate was washed
with 5 mL of PBS once or twice;
[0390] 3. PBS was sucked out. 1.5 mL of 0.25% pancreatin was added
to the plate to infiltrate the cells for 1 min;
[0391] 4. The culture plate was placed in an incubator. Digestion
was carried out for about 5 min at 37.degree. C.;
[0392] 5. 3 mL of complete culture solution was added to the
culture plate to stop the digestion. The cells were carefully
scoured with micropipette (1 mL) to give a uniform cell suspension.
The suspension was implanted into a 96-well cell culture disc by
2500 cells/100 .mu.L per well. The culture plate was incubated
overnight under 5% CO.sub.2 at 37.degree. C. In day 2, 100 .mu.L of
culture solution comprising a compound was added in each well. The
plate was further incubated for 72 h under 5% CO.sub.2 at
37.degree. C.;
[0393] 6. The culture solution was sucked out;
[0394] 7. 100 .mu.L of serum-free culture solution containing 0.5
mg/mL MTT was added in each well. The plate was incubated for 3
h;
[0395] 8. The culture solution was carefully sucked out;
[0396] 9. 100 .mu.L of DMSO was added in each well and vibrated to
dissolve;
[0397] 10. OD values were determined at 490 nM.
IV. Results and Treatments
[0398] 1. Calculation of Relative Inhibition Ratio
The inhibition ratio of a compound on cell
growth=(PC-n)/(PC-NC).times.100%
[0399] wherein:
[0400] PC: OD values of cells after normal growth in control wells
without a compound;
[0401] n: OD values of cells after growth in test wells with a
compound;
[0402] NC: Background OD values of blank wells without a compound
and cells.
TABLE-US-00010 TABLE 8 Growth Inhibition Activity of Some Compounds
in Examples on A431 Cells Inhibition Inhibition Inhibition Activity
Com- Activity Com- Activity Compounds (IC.sub.50/nM) pound
(IC.sub.50/nM) pound (IC.sub.50/nM) Compound 68 Example 42 Example
46 A 81 89 Example 97 45
Example 8
MCF-7 Cell Growth Inhibition Test (SRB Assay)
I. Assay Materials
[0403] Cell strains: MCF-7 (human breast cancer cell strains); SRB:
0.4% (w/v) working solution was formulated with 1% glacial acetic
acid, reserved at 4.degree. C.; antitumor compounds; compound A:
3'-pyrrolyldoxorubicin-14-oxo-succinic acid monoester; DMSO.
II. Reagents and Consumable Materials
[0404] Culture medium: 90% EMEM+10% FBS; Pancreatin (0.25% (w/v)
solution was formulated with PBS, 0.53 mM of EDTA was added in the
formulation); PBS; 96-well culture plate.
III. Assay Process
[0405] 1. A plate (10 cm) of cells in logarithmic growth phase that
were normally cultured was collected;
[0406] 2. The culture solution was sucked out. The plate was washed
with 5 mL of PBS once or twice;
[0407] 3. 1.5 mL of 0.25% pancreatin was added to the plate to
infiltrate the cells;
[0408] 4. The pancreatin was sucked out. The culture plate was
placed in an incubator. Digestion was carried out for about 3 min
at 37.degree. C.;
[0409] 5. 4 mL of complete culture solution was added to the
culture plate to stop the digestion. The cells were carefully
scoured with micropipette (1 mL) to give a uniform single cell
suspension. The suspension was implanted into a 96-well cell
culture plate by 10000 cells/100 .mu.L per well. The culture plate
was incubated overnight under 5% CO.sub.2 at 37.degree. C. In day
2, 100 .mu.L of culture solution comprising a compound was added in
each well. The plate was further incubated for 73 h under 5%
CO.sub.2 at 37.degree. C.;
[0410] 6. The culture solution was sucked out. 100 uL of TCA fixed
cells which were diluted to 10% were added to each well. The plate
was kept in a refrigerator for 1 h at 4.degree. C.
[0411] 7. TCA stationary liquid was sucked out. Each well was
washed with 150 .mu.L of ddH.sub.2O five times;
[0412] 8. After the stationary liquid was cleansed, the plate was
dried in the air at the room temperature;
[0413] 9. 60 .mu.L of SRB staining solution was added in each well.
The well was stained for 15 min at the room temperature;
[0414] 10. The SRB staining solution was sucked out. Each well was
washed with 150 .mu.L of 1% glacial acetic acid five times;
[0415] 11. After the SRB staining solution was cleansed, the plate
was dried in the air at the room temperature;
[0416] 12. 100 .mu.L of 10 mM Tris was added in each well. The
plate was vibrated to dissolve out SRB;
[0417] 13. OD values were determined at 570 nM.
IV. Results and Treatments
[0418] 1. Calculation of Relative Inhibition Ratio
The inhibition ratio of a compound on cell
growth=(PC-n)/(PC-NC).times.100%
[0419] wherein:
[0420] PC: OD values of cells after normal growth in control wells
without a compound;
[0421] n: OD values of cells after growth in test wells with a
compound;
[0422] NC: Background OD values of blank wells without a compound
and cells.
TABLE-US-00011 TABLE 9 Growth Inhibition Activity of Some Compounds
in Examples on MCF-7 Cells Inhibition Inhibition Inhibition
Activity Com- Activity Com- Activity Compound (IC.sub.50/nM) pound
(IC.sub.50/nM) pound (IC.sub.50/nM) Compound 785 Example 225
Example 333 A 81 89
Example 9
NCI-446 Cell Growth Inhibition Test (MTT Assay)
I. Assay Materials
[0423] Cell strains: NCI-446 (human small cell lung cancer cell
strains); MTT: nitroblue tetrazolium; antitumor compounds; compound
A: 3'-pyrrolyldoxorubicin-14-oxo-succinic acid monoester; DMSO.
II. Reagents and Consumable Materials
[0424] Culture medium: 90% RPMI1640+10% FBS (fetal calf serum);
Pancreatin (0.25% (w/v) solution was formulated with PBS, 0.53 mM
of EDTA was added in the formulation); PBS; 96-well culture
plate.
III. Assay Process
[0425] 1. A plate (10 cm) of cells in logarithmic growth phase that
were normally cultured was collected;
[0426] 2. The culture solution was sucked out. The plate was washed
with 5 ml of PBS once or twice;
[0427] 3. PBS was sucked out. 1.5 mL of 0.25% pancreatin was added
to the plate to infiltrate the cells for 1 min;
[0428] 4. The culture plate was placed in an incubator. Digestion
was carried out for about 3 min at 37.degree. C.;
[0429] 5. 3 mL of complete culture solution was added to the
culture plate to stop the digestion. The cells were carefully
scoured with micropipette (1 mL) to give a uniform cell suspension.
The suspension was implanted into a 96-well cell culture disc by
4000 cells/100 .mu.L per well. The culture plate was incubated
overnight under 5% CO.sub.2 at 37.degree. C. In day 2, 100 .mu.L of
culture solution comprising a compound was added in each well. The
plate was further incubated for 72 h under 5% CO.sub.2 at
37.degree. C.;
[0430] 6. The culture solution was sucked out;
[0431] 7. 100 .mu.L of serum-free culture solution containing 0.5
mg/mL MTT was added in each well. The plate was incubated for 3
h;
[0432] 8. The culture solution was carefully sucked out;
[0433] 9. 100 .mu.L of DMSO was added in each well and vibrated to
dissolve;
[0434] 10. OD values were determined at 490 nM.
IV. Results and Treatments
[0435] 1. Calculation of Relative Inhibition Ratio
The inhibition ratio of a compound on cell
growth=(PC-n)/(PC-NC).times.100%
[0436] wherein:
[0437] PC: OD values of cells after normal growth in control wells
without a compound;
[0438] n: OD values of cells after growth in test wells with a
compound;
[0439] NC: Background OD values of blank wells without a compound
and cells.
TABLE-US-00012 TABLE 10 Growth Inhibition Activity of Some
Compounds in Examples on NCI-446 Cells Inhibition Activity
Inhibition Activity Compound (IC.sub.50/nM) Compound (IC.sub.50/nM)
Compound A 234 Example 89 140
Example 10
NCI-H460 Cell Growth Inhibition Test (MTT Assay)
I. Assay Materials
[0440] Cell strains: NCI-H460 (human large cell lung cancer cell
strains); MTT: nitroblue tetrazolium; antitumor compounds; compound
A: 3'-pyrrolyldoxorubicin-14-oxo-succinic acid monoester; DMSO.
II. Reagents and Consumable Materials
[0441] Culture medium: 90% RPMI1640+10% FBS (fetal calf serum);
Pancreatin (0.25% (w/v) solution was formulated with PBS, 0.53 mM
of EDTA was added in the formulation); PBS; 96-well culture
plate.
III. Assay Process
[0442] 1. A plate (10 cm) of cells in logarithmic growth phase that
were normally cultured was collected;
[0443] 2. The culture solution was sucked out. The plate was washed
with 5 mL of PBS once or twice;
[0444] 3. PBS was sucked out. 1.5 mL of 0.25% pancreatin was added
to the plate to infiltrate the cells for 1 min;
[0445] 4. The culture plate was placed in an incubator. Digestion
was carried out for about 3 min at 37.degree. C.;
[0446] 5. 3 mL of complete culture solution was added to the
culture plate to stop the digestion. The cells were carefully
scoured with micropipette (1 mL) to give a uniform cell suspension.
The suspension was implanted into a 96-well cell culture disc by
2000 cells/100 .mu.L per well. The culture plate was incubated
overnight under 5% CO.sub.2 at 37.degree. C. In day 2, 100 .mu.L of
culture solution comprising a compound was added in each well. The
plate was further incubated for 72 h under 5% CO.sub.2 at
37.degree. C.;
[0447] 6. The culture solution was sucked out;
[0448] 7. 100 .mu.L of serum-free culture solution containing 0.5
mg/mL MTT was added in each well. The plate was incubated for 3
h;
[0449] 8. The culture solution was carefully sucked out;
[0450] 9. 100 .mu.L of DMSO was added in each well and vibrated to
dissolve;
[0451] 10. OD values were determined at 490 nM.
IV. Results and Treatments
[0452] 1. Calculation of Relative Inhibition Ratio
The inhibition ratio of a compound on cell
growth=(PC-n)/(PC-NC).times.100%
[0453] wherein:
[0454] PC: OD values of cells after normal growth in control wells
without a compound;
[0455] n: OD values of cells after growth in test wells with a
compound;
[0456] NC: Background OD values of blank wells without a compound
and cells.
TABLE-US-00013 TABLE 11 Growth Inhibition Activity of Some
Compounds in Examples on NCI-460 Cells Inhibition Activity
Inhibition Activity Compound (IC.sub.50/nM) Compound (IC.sub.50/nM)
Compound A 21 Example 89 2.5
Example 11
B16 Cell Growth Inhibition Test (MTT Assay)
I. Assay Materials
[0457] Cell strains: B16 (mouse melanoma cell strains); MTT:
nitroblue tetrazolium; antitumor compounds; compound A:
3'-pyrrolyldoxorubicin-14-oxo-succinic acid monoester; DMSO.
II. Reagents and Consumable Materials
[0458] Culture medium: 90% RPMI1640+10% FBS (fetal calf serum);
Pancreatin (0.25% (w/v) solution was formulated with PBS, 0.53 mM
of EDTA was added in the formulation); PBS; 96-well culture
plate.
III. Assay Process
[0459] 1. A plate (10 cm) of cells in logarithmic growth phase that
were normally cultured was collected;
[0460] 2. The culture solution was sucked out. The plate was washed
with 5 mL of PBS once or twice;
[0461] 3. PBS was sucked out. 1.5 mL of 0.25% pancreatin was added
to the plate to infiltrate the cells for 1 min;
[0462] 4. The culture plate was placed in an incubator. Digestion
was carried out for about 1 min at 37.degree. C.;
[0463] 5. 3 mL of complete culture solution was added to the
culture plate to stop the digestion. The cells were carefully
scoured with micropipette (1 mL) to give a uniform cell suspension.
The suspension was implanted into a 96-well cell culture disc by
2500 cells/100 .mu.L per well. The culture plate was incubated
overnight under 5% CO.sub.2 at 37.degree. C. In day 2, 100 .mu.L of
culture solution comprising a compound was added in each well. The
plate was further incubated for 72 h under 5% CO.sub.2 at
37.degree. C.;
[0464] 6. The culture solution was sucked out;
[0465] 7. 100 .mu.l of serum-free culture solution containing 0.5
mg/mL MTT was added in each well. The plate was incubated for 3
h;
[0466] 8. The culture solution was carefully sucked out;
[0467] 9. 100 .mu.L of DMSO was added in each well and vibrated to
dissolve;
[0468] 10. OD values were determined at 490 nM.
IV. Results and Treatments
[0469] 1. Calculation of Relative Inhibition Ratio
The inhibition ratio of a compound on cell
growth=(PC-n)/(PC-NC).times.100%
[0470] wherein:
[0471] PC: OD values of cells after normal growth in control wells
without a compound;
[0472] n: OD values of cells after growth in test wells with a
compound;
[0473] NC: Background OD values of blank wells without a compound
and cells.
TABLE-US-00014 TABLE 12 Growth Inhibition Activity of Some
Compounds in Examples on B16 Cells Inhibition Inhibition Inhibition
Activity Com- Activity Com- Activity Compound (IC.sub.50/nM) pound
(IC.sub.50/nM) pound (IC.sub.50/nM) Compound 32 Example 3 Example 3
A 62 89
Example 12
786-0 Cell Growth Inhibition Test (MTT Assay)
I. Assay Materials
[0474] Cell strains: 786-0 (human clear cell adenocarcinoma cell
strains); MTT: nitroblue tetrazolium; antitumor compounds; compound
A: 3'-pyrrolyldoxorubicin-14-oxo-succinic acid monoester; DMSO.
II. Reagents and Consumable Materials
[0475] Culture medium: 90% RPMI1640+10% FBS (fetal calf serum);
Pancreatin (0.25% (w/v) solution was formulated with PBS, 0.53 mM
of EDTA was added in the formulation); PBS; 96-well culture
plate.
III. Assay Process
[0476] 1. A plate (10 cm) of cells in logarithmic growth phase that
were normally cultured was collected;
[0477] 2. The culture solution was sucked out. The plate was washed
with 5 mL of PBS once or twice;
[0478] 3. PBS was sucked out. 1.5 mL of 0.25% pancreatin was added
to the plate to infiltrate the cells for 1 min;
[0479] 4. The culture plate was placed in an incubator. Digestion
was carried out for about 1 min at 37.degree. C.;
[0480] 5. 3 mL of complete culture solution was added to the
culture plate to stop the digestion. The cells were carefully
scoured with micropipette (1 mL) to give a uniform cell suspension.
The suspension was implanted into a 96-well cell culture disc by
2000 cells/100 .mu.L per well. The culture plate was incubated
overnight under 5% CO.sub.2 at 37.degree. C. In day 2, 100 .mu.L of
culture solution comprising a compound was added in each well. The
plate was further incubated for 72 h under 5% CO.sub.2 at
37.degree. C.;
[0481] 6. The culture solution was sucked out;
[0482] 7. 100 .mu.L of serum-free culture solution containing 0.5
mg/mL MTT was added in each well. The plate was incubated for 3
h;
[0483] 8. The culture solution was carefully sucked out;
[0484] 9. 100 .mu.L of DMSO was added in each well and vibrated to
dissolve;
[0485] 10. OD values were determined at 490 nM.
IV. Results and Treatments
[0486] 1. Calculation of Relative Inhibition Ratio
The inhibition ratio of a compound on cell
growth=(PC-n)/(PC-NC).times.100%
[0487] wherein:
[0488] PC: OD values of cells after normal growth in control wells
without a compound;
[0489] n: OD values of cells after growth in test wells with a
compound;
[0490] NC: Background OD values of blank wells without a compound
and cells.
TABLE-US-00015 TABLE 13 Growth Inhibition Activity of Some
Compounds in Examples on 786-O Cells Inhibition Inhibition
Inhibition Activity Com- Activity Com- Activity Compound
(IC.sub.50/nM) pound (IC.sub.50/nM) pound (IC.sub.50/nM) Compuond
180 Example 110 Example 90 A 62 90
Example 13
Du-145 Cell Growth Inhibition Test (MTT Assay)
I. Assay Materials
[0491] Cell strains: DU-145 (prostate cancer cell strains); MTT:
nitroblue tetrazolium; antitumor compounds; compound A:
3'-pyrrolyldoxorubicin-14-oxo-succinic acid monoester; DMSO.
II. Reagents and Consumable Materials
[0492] Culture medium: 90% EMEM+10% FBS (fetal calf serum);
Pancreatin (0.25% (w/v) solution was formulated with PBS, 0.53 mM
of EDTA was added in the formulation); PBS; 96-well culture
plate.
III. Assay Process
[0493] 1. A plate (10 cm) of cells in logarithmic growth phase that
were normally cultured was collected;
[0494] 2. The culture solution was sucked out. The plate was washed
with 5 mL of PBS once or twice;
[0495] 3. PBS was sucked out. 1.5 mL of 0.25% pancreatin was added
to the plate to infiltrate the cells for 1 min;
[0496] 4. The culture plate was placed in an incubator. Digestion
was carried out for about 1 min at 37.degree. C.;
[0497] 5. 3 mL of complete culture solution was added to the
culture plate to stop the digestion. The cells were carefully
scoured with micropipette (1 mL) to give a uniform cell suspension.
The suspension was implanted into a 96-well cell culture disc by
4000 cells/100 .mu.L per well. The culture plate was incubated
overnight under 5% CO.sub.2 at 37.degree. C. In day 2, 100 .mu.L of
culture solution comprising a compound was added in each well. The
plate was further incubated for 72 h under 5% CO.sub.2 at
37.degree. C.;
[0498] 6. The culture solution was sucked out;
[0499] 7. 100 .mu.L of serum-free culture solution containing 0.5
mg/mL MTT was added in each well. The plate was incubated for 3
h;
[0500] 8. The culture solution was carefully sucked out;
[0501] 9. 100 .mu.L of DMSO was added in each well and vibrated to
dissolve;
[0502] 10. OD values were determined at 490 nM.
IV. Results and Treatments
[0503] 1. Calculation of Relative Inhibition Ratio
The inhibition ratio of a compound on cell
growth=(PC-n)/(PC-NC).times.100%
[0504] wherein:
[0505] PC: OD values of cells after normal growth in control wells
without a compound;
[0506] n: OD values of cells after growth in test wells with a
compound;
[0507] NC: Background OD values of blank wells without a compound
and cells.
TABLE-US-00016 TABLE 14 Growth Inhibition Activity of Some
Compounds in Examples on DU-145 Cells Inhibition Inhibition
Inhibition Activity Com- Activity Com- Activity Compounds
(IC.sub.50/nM) pounds (IC.sub.50/nM) pound (IC.sub.50/nM) Compound
99 Example 34 Example 56 A 62 89 Compound 30 Example 61 90 92
Example 14
Hep3B Cell Growth Inhibition Test (SRB Assay)
I. Assay Materials
[0508] Cell strains: Hep3B (liver cancer cell strains); SRB: 0.4%
(w/v) working solution was formulated with 1% glacial acetic acid,
reserved at 4.degree. C.; antitumor compounds; compound A:
3'-pyrrolyldoxorubicin-14-oxo-succinic acid monoester; DMSO.
II. Reagents and Consumable Materials
[0509] Culture medium: 90% RPMI-1640+10% FBS; Pancreatin (0.25%
(w/v) solution was formulated with PBS, 0.53 mM of EDTA was added
in the formulation); PBS; 96-well culture plate.
III. Assay Process
[0510] 1. A plate (10 cm) of cells in logarithmic growth phase that
were normally cultured was collected;
[0511] 2. The culture solution was sucked out. The plate was washed
with 5 mL of PBS once or twice;
[0512] 3. 1.5 mL of 0.25% pancreatin was added to the plate to
infiltrate the cells;
[0513] 4. The pancreatin was sucked out. The culture plate was
placed in an incubator. Digestion was carried out for about 3 min
at 37.degree. C.;
[0514] 5. 4 mL of complete culture solution was added to the
culture plate to stop the digestion. The cells were carefully
scoured with micropipette (1 mL) to give a uniform single cell
suspension. The suspension was implanted into a 96-well cell
culture plate by 5000 cells/100 .mu.L per well. The culture plate
was incubated overnight under 5% CO.sub.2 at 37.degree. C. In day
2, 100 .mu.L of culture solution comprising a compound was added in
each well. The plate was further incubated for 72 h under 5%
CO.sub.2 at 37.degree. C.;
[0515] 6. The culture solution was sucked out. 100 uL of TCA fixed
cells which were diluted to 10% were added to each well. The plate
was kept in a refrigerator for 1 h at 4.degree. C.
[0516] 7. TCA stationary liquid was sucked out. Eeach well was
washed with 150 .mu.L of ddH.sub.2O five times;
[0517] 8. After the stationary liquid was cleansed, the plate was
dried in the air at the room temperature;
[0518] 9. 60 .mu.L of SRB staining solution was added in each well.
The well was stained for 15 min at the room temperature;
[0519] 10. The SRB staining solution was sucked out. Each well was
washed with 150 .mu.L of 1% glacial acetic acid five times;
[0520] 11. After the SRB staining solution was cleansed, the plate
was dried in the air at the room temperature;
[0521] 12. 100 .mu.L of 10 mM Tris was added in each well. The
plate was vibrated to dissolve out SRB;
[0522] 13. OD values were determined at 570 nM.
IV. Results and Treatments
[0523] 1. Calculation of Relative Inhibition Ratio
The inhibition ratio of the compound on cell
growth=(PC-n)/(PC-NC).times.100%
[0524] wherein:
[0525] PC: OD values of cells after normal growth in control wells
without a compound;
[0526] n: OD values of cells after growth in test wells with a
compound;
[0527] NC: Background OD values of blank wells without a compound
and cells.
TABLE-US-00017 TABLE 15 Growth Inhibition Activity of Some
Compounds in Examples on Hep3B Cells Inhibition Inhibition
Inhibition Activity Com- Activity Com- Activity Compound
(IC.sub.50/nM) pound (IC.sub.50/nM) pound (IC.sub.50/nM) Compound
444 Example 29 Example 28 A 90 92
Example 15
SK-Br-3 Cell Growth Inhibition Test (SRB Assay)
I. Assay Materials
[0528] Cell strains: SK-Br-3 (human breast cancer cell strains);
SRB: 0.4% (w/v) working solution was formulated with 1% glacial
acetic acid, reserved at 4.degree. C.; antitumor compounds;
compound A: 3'-pyrrolyldoxorubicin-14-oxo-succinic acid monoester;
DMSO.
II. Reagents and Consumable Materials
[0529] Culture medium: 85% DMEM+15% FBS; pancreatin (0.25% (w/v)
solution was formulated with PBS, 0.53 mM of EDTA was added in the
formulation); PBS; 96-well culture plate.
III. Assay Process
[0530] 1. A plate (10 cm) of cells in logarithmic growth phase that
were normally cultured was collected;
[0531] 2. The culture solution was sucked out. The plate was washed
with 5 mL of PBS once or twice;
[0532] 3. 1.5 mL of 0.25% pancreatin was added to the plate to
infiltrate the cells;
[0533] 4. The pancreatin was sucked out. The culture plate was
placed in an incubator. Digestion was carried out for about 2 min
at 37.degree. C.;
[0534] 5. 4 mL of complete culture solution was added to the
culture plate to stop the digestion. The cells were carefully
scoured with micropipette (1 mL) to give a uniform single cell
suspension. The suspension was implanted into a 96-well cell
culture plate by 10000 cells/100 .mu.L per well. The culture plate
was incubated overnight under 5% CO.sub.2 at 37.degree. C. In day
2, 100 .mu.L of culture solution comprising a compound was added in
each well. The plate was further incubated for 72 h under 5%
CO.sub.2 at 37.degree. C.;
[0535] 6. The culture solution was sucked out. 100 uL of TCA fixed
cells which were diluted to 10% were added to each well. The plate
was kept in a refrigerator for 1 h at 4.degree. C.
[0536] 7. TCA stationary liquid was sucked out. Each well was
washed with 150 .mu.L of ddH.sub.2O five times;
[0537] 8. After the stationary liquid was cleansed, the plate was
dried in the air at the room temperature;
[0538] 9. 60 .mu.L of SRB staining solution was added in each well.
The well was stained for 15 min at the room temperature;
[0539] 10. The SRB staining solution was sucked out. Each well was
washed with 150 .mu.L of 1% glacial acetic acid five times;
[0540] 11. After the SRB staining solution was cleansed, the plate
was dried in the air at the room temperature;
[0541] 12. 100 .mu.L of 10 mM Tris was added in each well. The
plate was vibrated to dissolve out SRB;
[0542] 13. OD values were determined at 570 nM.
IV. Results and Treatments
[0543] 1. Calculation of Relative Inhibition Ratio
The inhibition ratio of a compound on cell
growth=(PC-n)/(PC-NC).times.100%
[0544] wherein:
[0545] PC: OD values of cells after normal growth in control wells
without a compound;
[0546] n: OD values of cells after growth in test wells with a
compound;
[0547] NC: Background OD values of blank wells without a compound
and cells.
TABLE-US-00018 TABLE 16 Growth Inhibition Activity of Some
Compounds in Examples on SK-Br-3 Cells Inhibition Activity
Inhibition Activity Compound (IC.sub.50/nM) Compound (IC.sub.50/nM)
Compound A 93 Example 98 15
Assay on MTD (Maxium Tolerated Dose) In Vivo of Mice
Assay on Maxium Tolerated Dose of Compound
[0548] Assay animals: mice of ICR strain with a weight of 18-22 g,
purchased from Beijing Vital River Laboratories Limited, License
SCXK (jing) 2007-0001.
[0549] Administration Regimen: each experimental animal was
administered with corresponding dosage of tested substance once by
injection of caudal vain every three days. Each dosage was
initially designed as 5 times of administration. The specific time
of administration depended on the conditions of the animals. It
should be carefully observed the behavior conditions of the animals
within 2 hours after administration. The animal's behavior was
observed every 4 hours in the day of administration. The survival
conditions and weight of animals were recorded every day and
observed for whether abnormality occurred on the body surface
thereof. The experimental animals were sacrificed on 14 day after
administration and dissected for observation.
1. Tested Substance 1: 3'-Pyrrolyldoxorubicin
[0550] 1) Administration Groups
[0551] I group: (20 mg/kg) group;
[0552] II group: (25 mg/kg) group;
[0553] III group: (30 mg/kg) group;
[0554] IV group: solvent control group;
[0555] There were four groups in total. Each group consisted of
five male mice and five female mice.
[0556] 2) Assay Results
[0557] (1) The effects on weights of experimental animals are shown
in FIG. 1.
[0558] (2) The effects on survival ratio of experimental animals
are shown in Table 17.
TABLE-US-00019 TABLE 17 Number of Number Total Animals per of Dead
Mortality Dosage Times of Dosage Group Animals Rate (mg/kg)
Administration (mg/kg) / / (%) 20 4 80 5/5 5/5 100 25 4 100 5/5 5/5
100 30 3 90 5/5 5/5 100 Control 4 0 5/5 0/0 0 Note: : male mouse; :
female mouse
[0559] 3) Assay Results and Discussions
[0560] The experimental animals in the solvent control group
exhibited normally increased weight without death. Death occurred
in succession in the experimental animals after administration with
20 mg/kg or 25 mg/kg four times. Death occurred in succession in
the experimental animals after administration with 30 mg/kg three
times. The mortality rate is 100%. The MTD of the tested compound 1
is less than 20 mg/kg (0.0338 mmol/kg) according to the q4d.times.5
administration regimen.
2. Tested Compound 2: The Compound of Example 96
[0561] 1) Administration Groups
[0562] I group: (40 mg/kg) group;
[0563] II group: (45 mg/kg) group;
[0564] III group: (50 mg/kg) group;
[0565] IV group: (55 mg/kg) group;
[0566] V group: (60 mg/kg) group;
[0567] There were five groups in total. Each group consisted of six
male mice and six female mice.
[0568] 2) Assay Results
[0569] (1) The effects on weights of experimental animals are shown
in FIG. 2.
[0570] (2) The effects on survival ratio of experimental animals
are shown in Table 18.
TABLE-US-00020 TABLE 18 Number of Number Total Animals per of Dead
Mortality Dosage Times of Dosage Group Animals Rate (mg/kg)
Administration (mg/kg) / / (%) 40 5 200 6/6 0/1 8.33 45 5 225 6/6
0/0 (1/2a) 0 50 5 250 6/6 0/0 (1/1a) 0 55 5 275 6/6 5/4 (5/3a) 25
60 5 300 6/6 3/4 53.85 Note: : male mouse; : female mouse
[0571] a represents that the experimental animal died immediately
after the injection of drug rather than death caused by
cytotoxicity. The animal which died immediately after
administration was not counted in the calculation of mortality rate
(mortality rate=the number of experimental animals which did not
die immediately after injection/(the number of animals in group-the
number of experimental animals died immediately).times.100%).
[0572] 3) Assay Results and Discussions
[0573] There was one dead animal in the 40 mg/kg dosage group. The
mortality rate is 8.33%. The dead animal was dissected for
observation of various organs. The organs did not exhibit obvious
abnormality. However, there was several bite marks on the surface
of the skin of the dead animal. This animal did not have
significant weight loss. Therefore, it is presumed that the death
is caused by cytotoxicity factor rather than drug. Two experimental
animals died immediately after the fifth administration in the 50
mg/kg dosage group. The two dead animals did not have significant
weight loss. It is presumed that the death is caused by
cytotoxicity factor rather than drug. The MTD of the tested
compound 2 is 50-55 mg/kg (0.0525-0.0578 mmol/kg) according to the
q4d.times.5 administration regimen.
3. Tested Substance 3: 3'-Pyrrolyldoxorubicin-14-Oxo-Succinic Acid
Monoester
[0574] 1) Administration Groups
[0575] I group: (30 mg/kg) group;
[0576] II group: (40 mg/kg) group;
[0577] III group: (50 mg/kg) group;
[0578] IV group: (60 mg/kg) group;
[0579] There were four groups in total. Each group consisted of
five male mice and five female mice.
[0580] 2) Assay Results
[0581] (1) The effects on weights of experimental animals are shown
in FIG. 3.
[0582] (2) The effects on survival ratio of experimental animals
are shown in Table 19.
TABLE-US-00021 TABLE 19 Number of Number Total Animals per of Dead
Mortality Dosage Times of Dosage Group Animals Rate (mg/kg)
Administration (mg/kg) / / (%) 30 5 150 5/5 0/1 10 40 5 200 5/5 3/2
50 50 5 250 5/5 5/5 100 60 5 300 5/5 5/5 100 Note: : male mouse; :
female mouse
[0583] 3) Assay Results and Discussions
[0584] The lethality rates of mice of ICR strain are 10%, 50%,
100%, 100% respectively, according to the administration regimens
of 30 mg/kg, 40 mg/kg, 50 mg/kg, 60 mg/k of compounds. One
experimental animal dried without significant weight loss in the
animals of 30 mg/kg group, indicating that this dead animal in 30
mg/kg group does not exclude the cause of animal's individual
difference. The MTD of the tested compound 3 is approximately 30
mg/kg (0.0433 mmol/kg) according to the q4d.times.5 administration
regimen.
[0585] The above general description regarding the invention
disclosed herein and the description of the specific embodiments
thereof cannot be construed as the limitation to the technical
solutions of the invention. One of ordinary skill in the art can
add, delete or combine the technical features disclosed in the
above general description and/or specific embodiments (including
Examples) to form other technical solutions within the invention
according to the disclosure herein without departing from the
constitutive elements of the invention.
* * * * *